Micropeptides and use of same for modulating gene expression

ABSTRACT

Process for detecting and identifying micropeptides (miPEPs) encoded by a nucleotide sequence contained in the sequence of the primary transcript of a microRNA and use thereof for modulating gene expression.

FIELD OF THE INVENTION

The present invention relates to micropeptides (peptides encoded bymicroRNAs or “miPEPs”) and use thereof for modulating gene expression.

BACKGROUND OF THE INVENTION

The microRNAs (miRNAs) are small non-coding RNAs, about 21 nucleotidesin length after maturation, which control expression of target genes atthe post-transcriptional level, by degrading the target mRNA or byinhibiting its translation. The miRNAs occur in plants and animals.

The target genes are often key genes in developmental processes. Forexample they encode transcription factors or proteins of the proteasome.

The regulation of expression of the miRNAs is very poorly understood,but it is known in particular that the latter involves, like most codinggenes, an RNA polymerase II: this enzyme produces a primary transcript,called “pri-miRNA”, which is then matured by a protein complex inparticular containing the Dicer type enzymes. This maturation leadsfirstly to the formation of a precursor of miRNA called “pre-miRNA”,having a stem-loop secondary structure containing the miRNA and itscomplementary sequence miRNA*. Then the precursor is matured, whichleads to formation of a shorter double-stranded RNA containing the miRNAand the miRNA*. The miRNA is then manipulated by the RISC complex, whichcleaves the mRNA of the target gene or inhibits its translation.

Moreover, it has been shown that the presence of introns in the primarytranscript of the microRNA increases the expression of the maturemicroRNA (Schwab et al., EMBO Rep., 14(7): 615-21, 2013). However, owingto experimental difficulties, the primary transcripts of microRNAs, orpri-miRNAs, have received very little study.

About 50% of eukaryotic genes have small open reading frames withintheir 5′UTR region (5′ UnTranslated Region) upstream of the codingsequence. These small open reading frames (or “uORFs” for upstream ORFs)may perform a role of translation regulator, mainly in cis, bymodulating the fixation and the rate of the ribosomes on the mRNA, butalso in trans by an as yet unknown mechanism, by means of peptidesencoded by said uORFs (Combier et al., Gene Dev, 22: 1549-1559, 2008).By definition, the uORFS are present upstream of coding genes.

Recently, small ORFs have also been discovered in long intergenicnon-coding RNAs (lincRNAs), the putative function of which, if itexists, is not known (Ingolia et al., Cell, 147(4): 789-802, 2011;Guttman & Rinn, Nature, 482(7385): 339-46, 2012).

However, no example has yet been reported concerning the existence ofORFs encoding peptides within non-coding microRNAs. Until now, themicroRNAs, and by extension their primary transcript, have always beenregarded, based on their particular mode of action, as non-codingregulatory RNAs that do not produce any peptide.

SUMMARY OF THE INVENTION

One of the aspects of the invention is to propose peptides capable ofmodulating the expression of microRNAs.

Another aspect of the invention is to propose a means for modulating theexpression of one or more target genes of a microRNA.

The present invention offers the advantage of allowing easier and moreeffective control of the expression of genes targeted by the microRNAs,through a means other than the microRNA.

The invention thus relates to a process for detecting and identifying amicropeptide (miPEP) encoded by a nucleotide sequence contained in thesequence of the primary transcript of a microRNA,

comprising:

-   -   a) a step of detecting an open reading frame from 12 to 303        nucleotides in length contained in the sequence of the primary        transcript of said microRNA, then    -   b) a step of comparison between:        -   the accumulation of said microRNA in a specified eukaryotic            cell expressing said microRNA,        -   in the presence of a peptide encoded by a nucleotide            sequence that is identical or degenerate relative to that of            said open reading frame, said peptide being present in the            cell independently of transcription of the primary            transcript of said microRNA, and        -   the accumulation of said microRNA in a eukaryotic cell of            the same type as the aforesaid specified eukaryotic cell            expressing said microRNA, in the absence of said peptide,

in which a modulation of the accumulation of said microRNA in thepresence of said peptide relative to the accumulation of said microRNAin the absence of said peptide indicates the existence of a micropeptideencoded by said open reading frame.

The invention relates in particular to a process for detecting andidentifying a micropeptide (miPEP) encoded by a nucleotide sequencecontained in the sequence of the primary transcript of a microRNA,

comprising:

-   -   a) a step of detecting an open reading frame from 15 to 303        nucleotides in length contained in the sequence of the primary        transcript of said microRNA, then    -   b) a step of comparison between:        -   the accumulation of said microRNA in a specified eukaryotic            cell expressing said microRNA,        -   in the presence of a peptide encoded by a nucleotide            sequence that is identical or degenerate relative to that of            said open reading frame, said peptide being present in the            cell independently of transcription of the primary            transcript of said microRNA, and        -   the accumulation of said microRNA in a eukaryotic cell of            the same type as the aforesaid specified eukaryotic cell            expressing said microRNA, in the absence of said peptide,

in which a modulation of the accumulation of said microRNA in thepresence of said peptide relative to the accumulation of said microRNAin the absence of said peptide indicates the existence of a micropeptideencoded by said open reading frame.

The invention relates in particular to a process for detecting andidentifying a micropeptide (miPEP) encoded by a nucleotide sequencecontained in the sequence of the primary transcript of a microRNA,

comprising:

-   -   a) a step of detecting an open reading frame from 15 to 303        nucleotides in length, or of 12 nucleotides in length, contained        in the sequence of the primary transcript of said microRNA, then    -   b) a step of comparison between:        -   the accumulation of said microRNA in a specified eukaryotic            cell expressing said microRNA,        -   in the presence of a peptide encoded by a nucleotide            sequence that is identical or degenerate relative to that of            said open reading frame, said peptide being present in the            cell independently of transcription of the primary            transcript of said microRNA, and        -   the accumulation of said microRNA in a eukaryotic cell of            the same type as the aforesaid specified eukaryotic cell            expressing said microRNA, in the absence of said peptide,

in which a modulation of the accumulation of said microRNA in thepresence of said peptide relative to the accumulation of said microRNAin the absence of said peptide indicates the existence of a micropeptideencoded by said open reading frame.

The invention relates in particular to a process for detecting andidentifying a micropeptide (miPEP) encoded by a nucleotide sequencecontained in the sequence of the primary transcript of a microRNA,

comprising:

-   -   a) a step of detecting an open reading frame of 12 nucleotides        in length contained in the sequence of the primary transcript of        said microRNA, then    -   b) a step of comparison between:        -   the accumulation of said microRNA in a specified eukaryotic            cell expressing said microRNA,        -   in the presence of a peptide encoded by a nucleotide            sequence that is identical or degenerate relative to that of            said open reading frame, said peptide being present in the            cell independently of transcription of the primary            transcript of said microRNA, and        -   the accumulation of said microRNA in a eukaryotic cell of            the same type as the aforesaid specified eukaryotic cell            expressing said microRNA,        -   in the absence of said peptide,

in which a modulation of the accumulation of said microRNA in thepresence of said peptide relative to the accumulation of said microRNAin the absence of said peptide indicates the existence of a micropeptideencoded by said open reading frame.

In a first step, the process for detecting and identifying amicropeptide therefore consists of detecting, on the primary transcriptof a microRNA, the existence of an open reading frame potentiallyencoding a peptide.

For its part, the second step makes it possible to characterize saidpeptide, i.e. to determine whether said peptide corresponds to a peptidereally produced in the cell, by searching for an effect of said peptideon the accumulation of said microRNA.

In order to demonstrate an effect of the peptide on the accumulation ofthe microRNA, a large quantity of peptide is introduced into a firstcell expressing said microRNA. The accumulation of the microRNA in thisfirst cell is then measured and compared with the accumulation of themicroRNA in a second cell identical to the first, but not containingsaid peptide.

Observation of a variation of the quantities of microRNA between thecells in the presence and in the absence of the peptide thus indicates(i) that there is a peptide encoded on the primary transcript of saidmicroRNA, (ii) that the sequence of this peptide is encoded by the openreading frame identified on the primary transcript of said microRNA, and(iii) that said peptide has an effect on the accumulation of saidmicroRNA.

The invention is therefore based on the unexpected double observationmade by the inventors that, on the one hand, there are open readingframes that are able to encode micropeptides present on the primarytranscripts of microRNAs, and on the other hand that said micropeptidesare capable of modulating the accumulation of said microRNAs.

In particular, the invention relates to a process for detecting andidentifying a micropeptide (miPEP) encoded by a nucleotide sequencecontained in the sequence of the primary transcript of a microRNA,

comprising:

-   -   a) a step of detecting an open reading frame from 15 to 303        nucleotides in length contained in the sequence of the primary        transcript of said microRNA, then    -   b) a step of comparison between:        -   the accumulation of said microRNA in a specified eukaryotic            cell expressing the primary transcript of said microRNA,        -   in the presence of a peptide encoded by a nucleotide            sequence that is identical or degenerate relative to that of            said open reading frame, said peptide being present in the            cell independently of transcription of the primary            transcript of said microRNA, and        -   the accumulation of said microRNA in a eukaryotic cell of            the same type as the aforesaid specified eukaryotic cell            expressing the primary transcript of said microRNA,        -   in the absence of said peptide,

in which a modulation of the accumulation of said microRNA in thepresence of said peptide relative to the accumulation of said microRNAin the absence of said peptide indicates the existence of a micropeptideencoded by said open reading frame.

In particular, the invention relates to a process for detecting andidentifying a micropeptide (miPEP) encoded by a nucleotide sequencecontained in the sequence of the primary transcript of a microRNA,

comprising:

-   -   a) a step of detecting an open reading frame from 15 to 303        nucleotides in length, or of 12 nucleotides in length, contained        in the sequence of the primary transcript of said microRNA, then    -   b) a step of comparison between:        -   the accumulation of said microRNA in a specified eukaryotic            cell expressing the primary transcript of said microRNA,        -   in the presence of a peptide encoded by a nucleotide            sequence that is identical or degenerate relative to that of            said open reading frame, said peptide being present in the            cell independently of transcription of the primary            transcript of said microRNA, and        -   the accumulation of said microRNA in a eukaryotic cell of            the same type as the aforesaid specified eukaryotic cell            expressing the primary transcript of said microRNA,        -   in the absence of said peptide,

in which a modulation of the accumulation of said microRNA in thepresence of said peptide relative to the accumulation of said microRNAin the absence of said peptide indicates the existence of a micropeptideencoded by said open reading frame.

In particular, the invention relates to a process for detecting andidentifying a micropeptide (miPEP) encoded by a nucleotide sequencecontained in the sequence of the primary transcript of a microRNA,

comprising:

-   -   a) a step of detecting an open reading frame of 12 nucleotides        in length contained in the sequence of the primary transcript of        said microRNA, then    -   b) a step of comparison between:        -   the accumulation of said microRNA in a specified eukaryotic            cell expressing the primary transcript of said microRNA,        -   in the presence of a peptide encoded by a nucleotide            sequence that is identical or degenerate relative to that of            said open reading frame, said peptide being present in the            cell independently of transcription of the primary            transcript of said microRNA, and        -   the accumulation of said microRNA in a eukaryotic cell of            the same type as the aforesaid specified eukaryotic cell            expressing the primary transcript of said microRNA,        -   in the absence of said peptide,

in which a modulation of the accumulation of said microRNA in thepresence of said peptide relative to the accumulation of said microRNAin the absence of said peptide indicates the existence of a micropeptideencoded by said open reading frame.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1. Effects of overexpression of MtmiR171b (miR171b identified inMedicago truncatula) on the expression of the HAM1 and HAM2 genes (A) oron the number of lateral roots (B) in M. truncatula.

(A) The y-axis indicates the relative expression of MtmiR171b (left-handcolumns), of HAM1 (middle columns) or of HAM2 (right-hand columns) in acontrol plant (white columns) or in a plant in which MtmiR171b isoverexpressed (black columns). The error bar corresponds to the standarderror of the mean (number of individuals=10). The overexpression ofMtmiR171b induces a decrease in the expression of the HAM1 and HAM2genes.

(B) The y-axis indicates the mean number of lateral roots observed in acontrol plant (white column) or in a plant in which MtmiR171b isoverexpressed (black column). The error bar corresponds to the standarderror of the mean (number of individuals=100). The overexpression ofMtmiR171b leads to a reduction in the number of lateral roots.

FIG. 2. Effects of overexpression of MtmiPEP171b1 on the expression ofMtmiR171b and of the HAM1 and HAM2 genes (A) or on the number of lateralroots (B) in M. truncatula.

(A) The y-axis indicates the relative expression of MtmiPEP171b1 (graphon left), miR171b (graph on right, left-hand columns), of HAM1(accession No. MtGI9-TC114268) (graph on right, middle columns) or ofHAM2 (accession No. MtGI9-TC120850) (graph on right, right-hand columns)in a control plant (white columns) or in a plant in which MtmiPEP171b1is overexpressed (black columns). The error bar corresponds to thestandard error of the mean (number of individuals=10). Theoverexpression of MtmiPEP171b1 induces an increase in the accumulationof MtmiR171b, as well as a decrease in the expression of the HAM1 andHAM2 genes.

(B) The y-axis indicates the mean number of lateral roots observed in acontrol plant (white column) or in a plant in which MtmiPEP171b1 isoverexpressed (black column). The error bar corresponds to the standarderror of the mean (number of individuals=100). The overexpression ofMtmiPEP171b1 leads to a reduction in the number of lateral roots.

FIG. 3. Effects of MtmiPEP171b1 on the expression of MtmiR171b and theHAM1 and HAM2 genes (A) and on the number of lateral roots (B) in M.truncatula.

(A) The y-axis indicates the relative expression of MtmiR171b (left-handcolumns), of HAM1 (middle columns) or of HAM2 (right-hand columns) in acontrol plant (white columns) or in a plant treated by watering oncedaily for 5 days with MtmiPEP171b1 at 0.01 μM (light grey columns), 0.1μM (dark grey columns) or 1 μM (black columns). The error barcorresponds to the standard error of the mean (number ofindividuals=10). Application of MtmiPEP171b1 at different concentrationsinduces an increase in the accumulation of MtmiR171b, as well as adecrease in the expression of the HAM1 and HAM2 genes.

(B) The y-axis indicates the mean number of lateral roots observed in acontrol plant (white column) or in a plant treated by watering withMtmiPEP171b1 at 0.1 μM once daily for 5 days (black column). The errorbar corresponds to the standard error of the mean (number ofindividuals=100). Application of MtmiPEP171b1 at 0.1 μM leads to areduction in the number of lateral roots.

(C) The y-axis indicates the relative expression of MtmiR171b (left-handcolumns), of HAM1 (middle columns) or of HAM2 (right-hand columns) in acontrol plant (white columns) or in a plant treated by watering oncedaily for 5 days with MtmiPEP171b1 at 0.01 μM (grey columns), 0.1 μM(dark grey columns) or 1 μM (black columns) or with 0.01 μM of ascramble peptide (LIVSHLYSEKFDCMRKILRI, SEQ ID NO: 428) (light greycolumns) the amino acid composition of which is identical to miPEP171bbut the sequence of which is different. The error bar corresponds to thestandard error of the mean (number of individuals=10).

FIG. 4. Effects of MtmiPEP171b1 on the expression of pre-MtmiR171b (A)and of MtmiR171b (B) in M. truncatula.

The y-axis indicates the relative expression of the precursors of thedifferent forms of the microRNA in control plants (left-hand column) orin plants treated by watering once daily for 5 days with MtmiPEP171b1 at0.01 μM, 0.1 μM or 1 μM (right-hand columns). The error bar correspondsto the standard error of the mean (number of individuals=200).Application of MtmiPEP171b1 at different concentrations leads to anincrease in the accumulation of pre-MtmiR171b (A) and of MtmiR171b (B).

FIG. 5. Effects of overexpression of MtmiPEP171b1 (A) and effects ofMtmiPEP171b1 (B) on the expression of different precursors of microRNAsin M. truncatula.

The y-axis indicates the ratio of the expression of the precursors ofmicroRNAs in plants overexpressing MtmiPEP171b1 to the expression ofthese same precursors in control roots (A), or the ratio of theexpression of the precursors of microRNAs in plants treated withMtmiPEP171b1 (0.1 μM) to the expression of these same precursors incontrol roots (B). The different precursors of microRNAs tested areindicated from left to right on the x-axis, namely pre-MtmiR171b (SEQ IDNO: 246), pre-MtmiR169 (SEQ ID NO: 359), pre-MtmiR169a (SEQ ID NO: 360),pre-MtmiR171a (SEQ ID NO: 361), pre-MtmiR171h (SEQ ID NO: 362),pre-MtmiR393a (SEQ ID NO: 363), pre-MtmiR393b (SEQ ID NO: 364),pre-MtmiR396a (SEQ ID NO: 365) and pre-MtmiR396b (SEQ ID NO: 366). Theerror bar corresponds to the standard error of the mean (number ofindividuals=10). It is noted that MtmiPEP171b1 only leads to an effecton the accumulation of MtmiR171b and not on the other miRNAs.

FIG. 6. Effects of translation of MtmiPEP171b1 on the expression ofMtmiR171b demonstrated in the model plant Nicotiana benthamiana. They-axis indicates the relative expression of MtmiR171b in tobacco plantstransformed in order to express pri-MtmiR171b (white column) or amutated pri-MtmiR171b in which the codon ATG has been replaced with ATT(black column). The mutated pri-MtmiR171b is therefore incapable ofproducing MtmiPEP171b1. The error bar corresponds to the standard errorof the mean (number of individuals=30). It is noted that the absence oftranslation of MtmiPEP171b1 leads to a marked decrease in theaccumulation of miR171b.

FIG. 7. Effects of overexpression of MtmiPEP171b1 on the expression ofpre-MtmiR171b demonstrated in the model plant Nicotiana benthamiana.

The y-axis indicates the relative expression of pre-MtmiR171b in tobaccoplants that have been transformed in order to express MtmiR171b(left-hand column), MtmiR171b and MtmiPEP171b1 (middle column), orMtmiR171b and a mutated version of MtmiORF171b in which the start codonATG has been replaced with ATT (right-hand column). The error barcorresponds to the standard error of the mean (number ofindividuals=30). It is noted that the expression of MtmiPEP171b1increases the expression of MtmiR171b, and this effect is dependent onthe translation of MtmiORF171b to MtmiPEP171b1.

FIG. 8. Effects of MtmiPEP171b1 on the expression of pre-MtmiR171bdemonstrated in the model plant Nicotiana benthamiana.

The y-axis indicates the relative expression of MtmiR171b in tobaccoplants transformed in order to express MtmiR171b onto which MtmiPEP171b1has been sprayed (0.1 μM) twice, 12 h and then 30 min before sampling(right-hand column) or not (left-hand column). The error bar correspondsto the standard error of the mean (number of individuals=6). The peptideMtmiPEP171b1 applied by spraying induces an increase in the accumulationof MtmiR171b.

FIG. 9. Effects of MtmiPEP171b1 on the expression of pri-miR171b (A),pre-MtmiR171b (B) and MtmiR171b (C) demonstrated in the model plantNicotiana benthamiana.

The y-axis indicates the relative expression of the precursors of thedifferent forms of the microRNA in tobacco plants modified in order toexpress MtmiR171b (left-hand column) or modified in order to expressMtmiR171b and overexpress MtmiPEP171b1 (right-hand columns, FIG. 9A) ortreated with 0.1 μM of miPEP171b1 (FIGS. 9B and C). The error barcorresponds to the standard error of the mean (number ofindividuals=30). The overexpression of MtmiPEP171b1 or application ofmiPEP171b1 increases the accumulation of pri-MtmiR171b (A),pre-MtmiR171b (B) and MtmiR171b (C).

FIG. 10. Localization of MtmiPEP171b1 in tobacco leaf cells that havebeen modified in order to express MtmiPEP171b1.

The photographs show tobacco leaf cells modified in order to express theprotein GFP alone (left panel) or the protein GFP fused to MtmiPEP171b1(right panel). These observations indicate that MtmiPEP171b1 islocalized in small nuclear bodies.

FIG. 11. Effects of the expression of AtmiPEP165a (identified inArabidopsis thaliana) on the expression of AtmiR165a (A), and of theexpression of AtmiPEP319a2 (identified in Arabidopsis thaliana) onAtmiR319a (B), demonstrated in the model plant of tobacco.

(A) The y-axis indicates the relative expression of AtmiR165a in tobaccoplants modified in order to express AtmiR165a (left-hand column) or toexpress AtmiR165a and AtmiPEP165a (right-hand column).

(B) The y-axis indicates the relative expression of AtmiR319a in tobaccoplants modified in order to express AtmiR319a (left-hand column) or inorder to express AtmiR319a and AtmiPEP319a (right-hand column).

The error bar corresponds to the standard error of the mean (number ofindividuals=30). In both cases, it is noted that the expression ofmiORF, and therefore the production of miPEP, leads to an increase inthe accumulation of pre-miRNA.

FIG. 12. Effects of treatment with AtmiPEP165a on root growth inArabidopsis thaliana.

The photograph shows two plants of the same age: a control plant (planton the left) and a plant treated with AtmiPEP165a (plant on the right).The treatment with AtmiPEP165a leads to a phenotype with greatlyaccelerated root growth in Arabidopsis thaliana. The graph shows theexpression of pre-miR165 in response to treatment with increasing dosesof AtmiPEP165a.

FIG. 13. Conservation of the sequence of miPEP8 identified inDrosophila.

The sequences of miPEP8 (SEQ ID NO: 104) were deduced from the sequencesof miORF8 (SEQ ID NO: 208) of 12 different Drosophila species and werealigned. A histogram shows the conservation of each amino acid betweenthe sequences of miORF8 in the 12 species analysed.

FIG. 14. Evolution of the mass (kDa) and isoelectric point (pI) ofmiPEP8 in the Drosophila species.

The y-axis on the left indicates the size of the miPEP8 (in kD). They-axis on the right indicates the isoelectric point of the miPEP. Thex-axis indicates the origin of the miPEP, i.e. the Drosophila species.It is noted that despite a significant change in their size (by morethan a factor of 3), the charge of the miPEPs is still very basic (>9.8)in the 12 species studied.

FIG. 15. Effect of the addition of sequences on the function of miPEP.

The tobacco leaves were transformed in order to overexpress miPEP171b.These graphs show that the addition of sequences (tag His, HA or GFP)does not alter the function of miPEP. The y-axis indicates the relativeexpression of pre-MtmiR171b in tobacco plants that have been transformedin order to express MtmiR171b (left-hand column), MtmiR171b andMtmiPEP171b1 with or without addition of protein tags (right-handcolumns). The error bar corresponds to the standard error of the mean(number of individuals=6). It is noted that the expression ofMtmiPEP171b1 increases the expression of MtmiR171b, and this effect isindependent of the presence of tags.

FIG. 16. Expression of MtmiPEP171b1 in the root system of Medicagotruncatula.

The roots of Medicago truncatula were transformed in order to expressfusions between GUS protein (in blue) and the promoter of miR17b (A, E),the ATG of miPEP171b1 (B, F), whole miPEP171b1 (C, G) or ATG2 (secondATG located on the precursor, after miPEP) (D, H). It is clear thatthere is expression of miRNA in the root tips (A) as well as the lateralroots (E). The transcriptional (B, F) and translational (C, G) fusionsshow an expression of miPEP171b in the same tissues, whereas the nextATG is not active (D, H).

FIG. 17. Expression of DmmiPEP8 in cells of Drosophila melanogaster

The cells of Drosphila melanogaster were transfected in order tooverexpress DmmiPEP8 (OE miPEP8) or miPEP8 of which the translationstart codons have been mutated (OE miPEP8 mut). The y-axis indicates therelative expression of Pre-miR8. The error bar corresponds to thestandard error of the mean (number of independent experiments=6). It isnoted that the expression of DmmiPEP8 increases the expression ofDmmiR8, and this effect is linked to the translation of the mRNA.

FIG. 18. Impact of DmmiPEP8 on accumulation of DmmiR8 in cells of D.melanogaster

The cells of Drosphila melanogaster were transfected in order tooverexpress wild-type DmmiR8 (OE miR8) or DmmiR8 the translation startcodons of which have been mutated (OE miR8 miPEP8 mut). The y-axisindicates the relative expression of Pre-miR8. The error bar correspondsto the standard error of the mean (number of independent experiments=2).It is noted that the presence of DmmiPEP8 increases the expression ofDmmiR8.

FIG. 19. Impact of HsmiPEP155 on accumulation of HsmiR155 in cells ofHomo sapiens

HeLa cells of Homo sapiens had been transfected in order to overexpressHsmiPEP155 (OE miPEP155). The y-axis indicates the relative expressionof Pre-miR155. The error bar corresponds to the standard error of themean (number of independent experiments=2). It is noted that theexpression of HsmiPEP155 increases the expression of HsmiR155.

FIG. 20. Effects of translation of MtmiPEP171b1 on the expression ofMtmiR171b demonstrated in the model plant Nicotiana benthamiana.

The y-axis indicates the relative expression of MtmiR171b in tobaccoplants transformed in order to express pri-miR171b (left-hand column), apri-miR171b in which the miORF171b has been deleted (middle column) or amutated pri-miR171b in which the codon ATG has been replaced with ATT(right-hand column). The mutated pri-miR171b is therefore incapable ofproducing miPEP171b1. The error bar corresponds to the standard error ofthe mean (number of individuals=30). It is noted that the absence oftranslation of miPEP171b1 leads to a marked decrease in the accumulationof miR171b.

FIG. 21. Effects of overexpression of MtmiPEP171b1 on the expression ofMtmiR171b demonstrated in the model plant Nicotiana benthamiana.

The y-axis indicates the relative expression of MtmiR171b in tobaccoplants that had been transformed with a vector allowing the expressionof miPEP171b and either a second empty vector (white column), or avector allowing the expression of mtmiPEP171b (left black column), or avector in which the codon ATG of the ORF encoding mtmiPEP171b has beenreplaced with ATT (middle black column), or a vector in which thenucleotide sequence of the ORF has been mutated without modifying theamino acid sequence of the translated peptide (miPEP encoded by adegenerate ORF) (right black column). The error bar corresponds to thestandard error of the mean (number of individuals=30). It is noted thatthe expression of MtmiPEP171b1 increases the expression of MtmiR171b,and this effect is dependent on the translation of MtmiORF171b toMtmiPEP171b1.

FIG. 22. Effects of AtmiPEP165a on accumulation of AtmiR165a and of itstarget genes (PHAVOLUTA: PHV, PHABOL USA: PHB and REVOLUTA: REV).

The y-axis indicates the relative expression of AtmiR165a, PHV, PHB andREV in roots of Arabidopsis thaliana treated with water (control) ordifferent concentrations of AtmiPEP165a (0.01 μM, 0.1 μM, 1 μM or 10μM). The error bar corresponds to the standard error of the mean (numberof individuals=10).

The treatment of plants with higher and higher concentrations ofAtmiPEP165a demonstrates a dose-dependent effect of the accumulation ofAtmiR165a and the negative regulation of its target genes as a functionof the quantity of AtmiPEP165a.

FIG. 23. Effects of treatment with AtmiPEP164a on the expression ofAtmiR164a in A. thaliana.

The photographs show the results of a Northern blot analysis of theaccumulation of AtmiR164a in roots treated with water (control,photograph on left) or with 0.1 μM of a synthetic peptide, having asequence identical to that of AtmiPEP164a, dissolved in water (0.1 μMmiPEP164a). The RNA U6 is used as loading control making it possible toquantify the quantity of AtmiR164a.

This experiment was repeated 4 times independently and led to similarresults.

Treatment of shoots of A. thaliana with 0.1 μM of miPEP164a leads to anincrease in the accumulation of miR164a.

FIG. 24. Effects of treatment with AtmiPEP164a on the growth ofArabidopsis thaliana.

The photographs show two plants (top views and side views) after 3 weeksof growth: a control plant watered with water (A), and a plant wateredwith a composition of 0.1 μM of synthetic peptide corresponding toAtmiPEP164a (B). Watering plants of Arabidopsis thaliana withAtmiPEP164a increases plant growth significantly.

FIG. 25. Effects of treatment with AtmiPEP165a on the expression ofAtmiR165a in A. thaliana.

The photographs show the results of a Northern blot analysis of theaccumulation of AtmiR165a in roots treated with water (control,photograph on left) or with 0.1 μM of a synthetic peptide, having asequence identical to that of AtmiPEP165a, dissolved in water (0.1 μMmiPEP165a). The RNA U6 is used as loading control making it possible toquantify the quantity of AtmiR165a.

This experiment was repeated 4 times independently and led to similarresults.

Treatment of A. thaliana shoots with 0.1 μM of miPEP165a leads to anincrease in the accumulation of miR165a.

FIG. 26. Effects of overexpression of AtmiPEP319a1 on the expression ofAtmiR319a in A. thaliana.

The y-axis indicates the relative expression of AtmiR319a in a controlplant (left-hand column) or in a plant in which AtmiPEP319a1 isoverexpressed (right-hand column). The error bar corresponds to thestandard error of the mean (number of individuals=10). Theoverexpression of AtmiPEP319a1 induces an increase in the accumulationof AtmiR319a.

FIG. 27. Effects of treatment with AtmiPEP319a on the growth ofArabidopsis thaliana.

The photographs show two plants (top views and side views) after 3 weeksof growth: a control plant watered with water (A), and a plant wateredwith a composition of 0.1 μM of synthetic peptide corresponding toAtmiPEP319a1 (B). Watering of the plants of Arabidopsis thaliana withAtmiPEP319a1 increases plant growth significantly.

FIG. 28. Immunolocalization.

The roots of Medicago truncatula were transformed in order to expressfusions between the GUS protein (in blue) and the ATG of miPEP171b(Pro_(miR171b)-ATG1:GUS) or ATG2 (second ATG located on the precursor,after miPEP) (Pro_(miR171b)-ATG2:GUS). Labelling was also carried outwith an anti-miPEP171b antibody (miPEP171b). Immunolocalization ofmiPEP171b in the roots of M. truncatula reveals the presence ofmiPEP171b in the lateral root initiation sites, showing aco-localization between the microRNA and the corresponding miPEP.

FIG. 29. Effects of the overexpression of AtmiPEP160b1 on the expressionof AtmiR160b in A. thaliana.

The y-axis indicates the relative expression of AtmiR160b in a controlplant (left-hand column) or in a plant in which AtmiPEP160b1 isoverexpressed (right-hand column). The error bar corresponds to thestandard error of the mean (number of individuals=10). Theoverexpression of AtmiPEP160b1 induces an increase in the relativeexpression of AtmiR160b.

FIG. 30. Effects of AtmiPEP164a1 on the expression of AtmiR164a.

The y-axis indicates the relative expression of AtmiR164a in a controlplant (left-hand column) or in a plant watered once a day withAtmiPEP164a1 at 0.1 μM (right-hand column). The error bar corresponds tothe standard error of the mean (number of individuals=10).

The addition of AtmiPEP164a1 induces induces an increase in the relativeexpression of AtmiR164a.

FIG. 31. Effects of the overexpression of AtmiPEP319a1 on the expressionof AtmiR319a in A. thaliana.

The y-axis indicates the relative expression of AtmiR319a in a controlplant (left-hand column) or in a plant in which AtmiPEP319a1 isoverexpressed (right-hand column). The error bar corresponds to thestandard error of the mean (number of individuals=10).

The overexpression of AtmiPEP319a1 induces an increase in the relativeexpression of AtmiR319a.

FIG. 32. Effects of MtmiPEP169d on the expression of MtmiR169d in M.truncatula.

The y-axis indicates the relative expression of MtmiR169d in a controlplant (left-hand column) or in a plant watered once a day withMtmiPEP169d at 0.1 μM (right-hand column).

The error bar corresponds to the standard error of the mean (number ofindividuals=10).

The addition of MtmiPEP169d induces an increase in the relativeexpression of MtmiR169d.

FIG. 33. Effects of the overexpression of MtmiPEP171e on the expressionof MtmiR171e in M. truncatula.

The y-axis indicates the relative expression of MtmiR171e in a controlplant (left-hand column) or in a plant in which MtmiPEP171e isoverexpressed (right-hand column). The error bar corresponds to thestandard error of the mean (number of individuals=10).

The overexpression of MtmiPEP171e induces an increase in the relativeexpression of MtmiR171e.

FIG. 34. Localization of the MtmiPEP171b1 in a wildtype plant M.truncatula.

Various amounts of synthetic peptides MtmiPEP171b1 (1, 0.5, or 0.1 nmol)and a total root extract from M. truncatula have been analysed byimmunoblotting with an antibody specific for MtmiPEP171b1.

MtmiPEP171b1 is naturally produced in the roots of M. truncatula.

FIG. 35. Presence of AtmiPEP165a in a wildtype plant A. thaliana.

The upper picture corresponds to the western blotting analysis of theamount of AtmiPEP165a in a wildtype seeding plant of A. thaliana Col-0(left-hand) and a seeding plant of A. thaliana overexpressingAtmiPEP165a (right-hand). The lower picture corresponds to the westernblotting analysis of the amount of a control protein (tubulin) in awildtype seeding plant of A. thaliana Col-0 (left-hand) and a seedingplant of A. thaliana overexpressing AtmiPEP165a (right-hand).AtmiPEP165ais naturally produced in A. thaliana and is present in larger amounts inplants overexpressing AtmiPEP165a.

FIG. 36. Action of AtmiPEP165a in A. thaliana.

(A) The y-axis indicates the relative expression of pri-miR165a incontrol roots (left-hand column) or watered once a day with AtmiPEP165aat 10 μM (right-hand column). The error bar corresponds to the standarderror of the mean (number of individuals=10).

The addition of AtmiPEP165a induces an increase in the relativeexpression of AtmiR165a.

(B) The y-axis indicates the relative expression of pri-miR165a, in thepresence of cordycepin (inhitor of the RNA synthesis), in control roots(grey curve) or watered once a day with with AtmiPEP165a at 10 μM (blackcurve). The x-axis indicates the duration of the treatment withcordycepin.

The addition of AtmiPEP165a does not induce a better stability ofpri-miR165a.

(C) The y-axis indicates the relative expression of pri-miR165a, inwildtype plants (Col-0) or in plants mutated with a low allele of thesecond large subunit of RNA polymerase II (nrpb2-3). The x-axisindicates if the plants have been watered with water or with AtmiPEP165aat 10 at 10 μM.

Mutated plants, unlike wildtype plants, are not capable of accumulatingAtmiR165a in response to AtmiPEP165a.

FIG. 37. Effects of the pri-miR171b on the number of lateral roots in M.truncatula.

The y-axis indicates the mean number of lateral roots observed in acontrol plant (white column) or in a plant in which the pri-miR171b isoverexpressed (black column). The error bar corresponds to the standarderror of the mean (number of individuals=100). The overexpression ofpri-miR171b induces a decrease in the number of lateral roots.

FIG. 38. Translatability of DmmiPEP8 in Drosophila melanogaster cells

Some Drosophila melanogaster cells have been transfected to overexpressDmmiPEP8 (miPEP8::GFP) or DmmiPEPmt (of which start codons have beenmutated (miPEP8 mt::GFP). As a transfection control, a plasmid producingmoesin-RFP protein (lower picture) has been co-transfected with theplasmids encoding DmmiPEP8 (left-hand picture) and DmmiPEP8 mt(right-hand picture). It is observed that the start codons of theDmmiPEP8 sequence are functional because they allow the synthesis of GFP(top left picture) whereas the constructions with mutated ATGs do notproduce hardly any GFP (top right picture).

Thus, these results suggest that the ORF of the miPEP is functional.

DETAILED DESCRIPTION OF THE INVENTION

In the invention, the terms “microRNA”, “non-coding microRNA” and“miRNA” are equivalent and may be used interchangeably. They definesmall molecules of RNA of about 21 nucleotides, which are not translatedand do not lead to a peptide or a protein.

However, in this mature form, the microRNAs perform a function ofregulation of certain genes via post-transcriptional mechanisms, forexample by means of the RISC complex.

The primary transcript of the microRNA or “pri-miRNA” corresponds to theRNA molecule obtained directly from transcription of the DNA molecule.Generally, this primary transcript undergoes one or morepost-transcriptional modifications, involving for example a particularstructure of the RNA or cleavage of certain portions of the RNA bysplicing phenomena, and which lead to the precursor form of the microRNAor “pre-miRNA”, then to the mature form of the microRNA or “miRNA”.

The terms “micropeptides” and “miPEPs” (microRNA encoded PEPtides) areequivalent and may be used interchangeably. They define a peptide thatis encoded by an open reading frame present on the primary transcript ofa microRNA, and which is capable of modulating the accumulation of saidmicroRNA.

The micropeptides within the meaning of the present invention are not tobe understood as necessarily being small peptides, as “micro” does notcorrespond to the size of the peptide. According to the invention, amiPEP can also be considered as a transcription modulator, and inparticular a transcription activator. Such a transcription modulator canoperate at the transcription level to modulate the accumulation ofpri-miR, pre-miR and miR.

Taking into account the degeneracy of the genetic code, one and the samemicropeptide may be encoded by several nucleotide sequences. Nucleotidesequences of this kind, differing from one another by at least onenucleotide but encoding one and the same peptide, are called “degeneratesequences”.

The terms “open reading frame” or “ORF” are equivalent and may be usedinterchangeably. They correspond to a nucleotide sequence in a DNA orRNA molecule that may potentially encode a peptide or a protein: saidopen reading frame begins with a start codon (the start codon generallyencoding a methionine), followed by a series of codons (each codonencoding an amino acid), and ends with a stop codon (the stop codon notbeing translated).

In the invention, the ORFs may be called specifically “miORFs” when theyare present on the primary transcripts of microRNA.

The miORFs as defined in the particular invention may have a size from12 to 303 nucleotides and may encode peptides from 3 to 100 amino acids.

In particular, the miORFs as defined in the invention may have a sizefrom 15 to 303 nucleotides. As an amino acid is encoded by a codon of 3nucleotides, the miORFs from 15 to 303 nucleotides encode miPEPS from 4to 100 amino acids.

In particular, the miORFs have a size of:

15, 18, 21, 24, 27, 30, 33, 36, 39, 42, 45, 47, 51, 54, 57, 60, 63, 66,69, 72, 75, 78, 81, 84, 87, 90, 93, 96, 99, 102, 105, 108, 111, 114,117, 120, 123, 126, 129, 132, 135, 138, 141, 15 144, 147, 150, 153, 156,159, 162, 165, 168, 171, 174, 177, 180, 183, 186, 189, 192, 195, 198,201, 204, 207, 210, 213, 216, 219, 222, 225, 228, 231, 234, 237, 240,243, 246, 249, 252, 255, 258, 261, 264, 267, 270, 273, 276, 279, 282,285, 288, 291, 294, 297, 300 or 303 nucleotides, and encode respectivelymiPEPs having a size of:

4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22,23, 24, 25, 26, 27, 28, 29, 20 30, 31, 32, 33, 34, 35, 36, 37, 38, 39,40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57,58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75,76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93,94, 95, 96, 97, 98, 99 or 100 amino acids.

In the invention, “accumulation” means the production of a molecule,such as a microRNA or a micropeptide, in the cell.

Thus, “modulation” of the accumulation of a molecule in a cellcorresponds to a modification of the quantity of this molecule presentin the cell.

Moreover, the effect of a miPEP can be observed through the modulationof the accumulation of the miR, but also through the modulation of theaccumulation of the corresponding pri-miR or pre-miR.

In an embodiment, the invention relates to a process for detecting andidentifying a miPEP as defined above, in which the modulation of theaccumulation of said microRNA is a decrease or an increase in theaccumulation of said microRNA, in particular an increase.

A “decrease in the accumulation” corresponds to a decrease in thequantity of said molecule in the cell.

Conversely, an “increase in the accumulation” corresponds to an increasein the quantity of said molecule in the cell.

In an advantageous embodiment, the invention relates to a process fordetecting and identifying a miPEP as defined above, in which themodulation of the accumulation of said microRNA is an increase in theaccumulation of said microRNA.

In an embodiment, the invention relates to a process for detecting andidentifying a miPEP as defined above, in which the presence of saidpeptide in the cell results from:

-   -   the introduction of a nucleic acid encoding said peptide into        the cell, or    -   the introduction of said peptide into the cell.

In order to characterize a miPEP, it is necessary to have a cellularmodel expressing a microRNA in which said peptide to be tested ispresent. For this, it is possible to introduce a peptide into the cell,either by bringing the cell into contact with said peptide, or byintroducing a nucleic acid encoding said peptide into the cell, and thisnucleic acid will then be translated into peptide within the cell.

In an embodiment, the invention relates to a process for detecting andidentifying a miPEP as defined above, in which said open reading framein step a) is contained in the 5′ or 3′ portion of said primarytranscript of the microRNA, preferably in the 5′ portion.

The 5′ or 3′ portions of the primary transcript of the microRNAcorrespond to the terminal portions of the RNA molecule that are cleavedduring maturation of the microRNA.

In an embodiment, the invention relates to a process for detecting andidentifying a miPEP as defined above, in which said microRNA is presentin a wild-type plant cell.

In the invention, a wild-type plant cell corresponds to a plant cellthat has not been genetically modified by humans.

In an embodiment, the invention relates to a process for detecting andidentifying a miPEP as defined above, in which said microRNA is presentin a wild-type animal cell, and in particular a wild-type human cell ora wild-type Drosophila cell.

In the invention, a wild-type animal cell corresponds to an animal cell,and in particular a human cell, that has not been modified geneticallyby humans.

In an embodiment, the invention relates to a process for detecting andidentifying a miPEP as defined above, in which said specified eukaryoticcell and said eukaryotic cell of the same type as the aforesaidspecified eukaryotic cell, used in step b, are plant cells of acruciferous plant such as Arabidopsis thaliana, of a leguminous plantsuch as Glycine max (soya), Medicago truncatula and Medicago sativa(alfalfa) or of a plant of the Solanaceae family such as Nicotianabenthamiana (tobacco), Solanum tuberosum (potato), Solanum lycopersicum(tomato) or Solanum melongena (aubergine).

In an embodiment, the invention relates to a process for detecting andidentifying a miPEP as defined above, in which said specified eukaryoticcell and said eukaryotic cell of the same type as the aforesaidspecified eukaryotic cell, used in step b, are plant cells, preferablycells of Medicago truncatula, Nicotiana benthamiana or Arabidopsisthaliana.

In an embodiment, the invention relates to a process for detecting andidentifying a miPEP as defined above, in which said specified eukaryoticcell and said eukaryotic cell of the same type as the aforesaidspecified eukaryotic cell, used in step b, are animal cells, preferablyhuman cells or Drosophila cells.

In the process for detecting and identifying a micropeptide as definedabove, after identifying an ORF that is able to encode a peptide on theprimary transcript of a microRNA, it is necessary to have a cellularmodel having said microRNA and said peptide, so as to be able todemonstrate a possible effect of the peptide on said microRNA. Twooptions are therefore conceivable:

-   -   the cellular model in which the miORF has been identified and        that in which the effect of the peptide on the miRNA has been        demonstrated are identical, or    -   the cellular model in which the miORF has been identified and        that in which the effect of the peptide on the miRNA has been        demonstrated are different.

In the first option, the cellular model used for observing an effect ofthe peptide is the same as that in which the primary transcript of saidmicroRNA was isolated. In this cellular model, the specified eukaryoticcells contain said microRNA naturally and only the peptide to be testedhas to be introduced into these cells. In this context, said microRNA isqualified as “of endogenous origin” as it exists naturally in the cells.Nevertheless, other copies of a microRNA of endogenous origin may beadded to a cell, for example by introducing a vector encoding saidmicroRNA of endogenous origin into the cell.

In the second option, the cellular model used for observing an effect ofthe peptide is different from that in which the primary transcript ofsaid microRNA was isolated. In this cellular model, the specifiedeukaryotic cells contain neither the microRNA, nor the peptide to betested. These two elements must therefore be introduced into thesecells. In this context, said microRNA is qualified as “of exogenousorigin” as it does not exist naturally in the cells.

In an embodiment, the invention relates to a process for detecting andidentifying a miPEP as defined above, in which said microRNA is ofendogenous origin in said eukaryotic cell and in said eukaryotic cell ofthe same type as the aforesaid specified eukaryotic cell, used in stepb).

In an embodiment, the invention relates to a process for detecting andidentifying a miPEP as defined above in which said microRNA is ofexogenous origin in said eukaryotic cell and in said eukaryotic cell ofthe same type as the aforesaid specified eukaryotic cell, used in stepb), said eukaryotic cells containing a vector allowing the expression ofsaid microRNA.

In an embodiment, the invention relates to a process for detecting andidentifying a miPEP as defined above, in which the accumulation of saidmicroRNA is determined using quantitative RT-PCR or Northern blot.

In an embodiment, the invention relates to a process for detecting andidentifying a miPEP as defined above, in which the accumulation of saidmicroRNA is determined using a DNA or RNA chip.

The accumulation of said microRNA may be determined using the techniquesof molecular biology for assaying specific nucleic acid molecules.

In another aspect, the invention also relates to a process for detectingand identifying a microRNA in which the sequence of the primarytranscript contains a nucleotide sequence encoding a miPEP, comprising:

-   -   a) a step of detecting an open reading frame from 15 to 303        nucleotides in length contained in the sequence of the primary        transcript of said microRNA, then    -   b) a step of comparison between:        -   the accumulation of said microRNA in a specified eukaryotic            cell expressing said microRNA,        -   in the presence of a peptide encoded by a nucleotide            sequence that is identical or degenerate relative to that of            said open reading frame, said peptide being present in the            cell independently of transcription of the primary            transcript of said microRNA, and        -   the accumulation of said microRNA in a eukaryotic cell, of            the same type as the aforesaid specified eukaryotic cell            expressing said microRNA,        -   in the absence of said peptide,

in which a modulation of the accumulation of said microRNA in thepresence of said peptide relative to the accumulation of said microRNAin the absence of said peptide indicates the existence of a microRNA theprimary transcript of which contains a nucleotide sequence encoding amicropeptide.

In particular, the invention relates to a process for detecting andidentifying a microRNA in which the sequence of the primary transcriptcontains a nucleotide sequence encoding a miPEP,

comprising:

-   -   a) a step of detecting an open reading frame from 15 to 303        nucleotides in length contained in the sequence of the primary        transcript of said microRNA, then    -   b) a step of comparison between:        -   the accumulation of said microRNA in a specified eukaryotic            cell expressing the primary transcript of said microRNA,        -   in the presence of a peptide encoded by a nucleotide            sequence that is identical or degenerate relative to that of            said open reading frame, said peptide being present in the            cell independently of transcription of the primary            transcript of said microRNA, and        -   the accumulation of said microRNA in a eukaryotic cell, of            the same type as the aforesaid specified eukaryotic cell            expressing the primary transcript of said microRNA,        -   in the absence of said peptide,

in which a modulation of the accumulation of said microRNA in thepresence of said peptide relative to the accumulation of said microRNAin the absence of said peptide indicates the existence of a microRNA theprimary transcript of which contains a nucleotide sequence encoding amicropeptide.

In an embodiment, the invention relates to a process for detecting andidentifying a microRNA as defined above, in which the modulation of theaccumulation of said microRNA is a decrease or an increase in theaccumulation of said microRNA, in particular an increase.

In an embodiment, the invention relates to a process for detecting andidentifying a microRNA as defined above, in which the presence of saidpeptide in the cell results from:

-   -   the introduction of a nucleic acid encoding said peptide into        the cell, or    -   the introduction of said peptide into the cell.

In an embodiment, the invention relates to a process for detecting andidentifying a microRNA as defined above, in which said open readingframe in step a) is contained in the 5′ or 3′ portion of said primarytranscript of the microRNA, preferably in the 5′ portion. In anembodiment, the invention relates to a process for detecting andidentifying a microRNA as defined above, in which said microRNA ispresent in a wild-type plant cell.

In an embodiment, the invention relates to a process for detecting andidentifying a microRNA as defined above, in which said microRNA ispresent in a wild-type animal cell, and in particular a wild-type humancell.

In an embodiment, the invention relates to a process for detecting andidentifying a microRNA as defined above, in which said eukaryotic cell,and said eukaryotic cell of the same type as the aforesaid specifiedeukaryotic cell, used in step b) are plant cells, preferably cells ofMedicago truncatula.

In an embodiment, the invention relates to a process for detecting andidentifying a microRNA as defined above, in which said eukaryotic cell,and said eukaryotic cell of the same type as the aforesaid specifiedeukaryotic cell, used in step b) are animal cells, preferably Drosophilacells.

In an embodiment, the invention relates to a process for detecting andidentifying a microRNA as defined above, in which said microRNA is ofendogenous origin in said eukaryotic cell and in said eukaryotic cell ofthe same type as the aforesaid specified eukaryotic cell, used in stepb).

In an embodiment, the invention relates to a process for detecting andidentifying a microRNA as defined above in which said microRNA is ofexogenous origin in said eukaryotic cell and in said eukaryotic cell ofthe same type as the aforesaid specified eukaryotic cell, used in stepb), said eukaryotic cells containing a vector allowing the expression ofsaid microRNA.

In an embodiment, the invention relates to a process for detecting andidentifying a microRNA as defined above, in which the accumulation ofsaid microRNA is determined using quantitative RT-PCR or Northern blot.

In an embodiment, the invention relates to a process for detecting andidentifying a microRNA as defined above, in which the accumulation ofsaid microRNA is determined using a DNA or RNA chip.

In another aspect, the invention relates to a miPEP as obtained byimplementing the process as defined above.

More particularly, the invention relates to a miPEP encoded by anucleotide sequence as obtained by implementing the process as definedabove. In other words, the invention relates to a miPEP encoded by anucleotide sequence detected and identified by implementing the processas defined above.

Another aspect of the invention also relates to a miPEP of 3 to 100amino acids, in particular of 4 to 100 amino acids, in particular of 4to 60 amino acids, preferably of 4 to 40 amino acids, encoded by anucleotide sequence contained in the primary transcript of a microRNA,said miPEP being capable of modulating the accumulation of said microRNAin a eukaryotic cell.

Another aspect of the invention also relates to a miPEP of 4 to 100amino acids, or of 3 amino acids, encoded by a nucleotide sequencecontained in the primary transcript of a microRNA, said miPEP beingcapable of modulating the accumulation of said microRNA in a eukaryoticcell.

Another aspect of the invention also relates to a miPEP of 3 amino acidsencoded by a nucleotide sequence contained in the primary transcript ofa microRNA, said miPEP being capable of modulating the accumulation ofsaid microRNA in a eukaryotic cell.

In particular, the miPEP as defined in the invention is encoded by amiORF of 15 to 303 nucleotides and has a size of 4, 5, 6, 7, 8, 9, 10,11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28,29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 2546, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63,64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81,82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99or 100 amino acids, in particular 5, 8, 10, 18, 19, 23, 37, 50 or 59amino acids.

In particular, the miPEP of the invention has a size in the range from 4to 10 amino acids, 4 to 20 amino acids, 4 to 30 amino acids, 4 to 40amino acids, 4 to 50 amino acids, 4 to 60 amino acids, 4 to 70 aminoacids, 4 to 80 amino acids, 4 to 90 amino acids, or 4 to 100 aminoacids.

Moreover, it should be noted that several miORFS may be identified onthe primary transcript of a microRNA, indicating that a primarytranscript of microRNA may potentially encode several miPEPs.

It should also be noted that the effect of a miPEP is generally specificto a single microRNA, namely that resulting from the primary transcriptencoding said miPEP.

The modulation of the microRNA by said miPEP may be demonstrated afterobserving a variation in quantities of microRNA between the cells in thepresence and in the absence of the miPEP.

In an embodiment, the invention relates to a miPEP as defined above,said nucleotide sequence being contained in the 5′ or 3′ portion of saidprimary transcript of a microRNA, preferably in the 5′ portion.

In an embodiment, the invention relates to a miPEP as defined above,said nucleotide sequence corresponding to the first open reading framepresent on said primary transcript of a microRNA.

In an embodiment, the invention relates to a miPEP as defined above,said miPEP having a basic isoelectric point, preferably above 8.

In an embodiment, the invention relates to a miPEP as defined above,said miPEP having an acidic isoelectric point.

In an embodiment, the invention relates to a miPEP as defined above,said miPEP being selected from the group of peptides consisting of SEQID NO: 1 to SEQ ID NO: 104, SEQ ID NO: 375 to SEQ ID NO: 386, and SEQ IDNO: 355 (Table 1).

In an embodiment, the invention relates to a miPEP as defined above,said miPEP being selected from the group of peptides consisting of:

-   -   SEQ ID NO: 1 to SEQ ID NO: 104 and SEQ ID NO: 355,    -   SEQ ID NO: 375 to SEQ ID NO: 386, or    -   SEQ ID NO: 424.

In an embodiment, the invention relates to a miPEP as defined above,consisting of the amino acid sequence MVT.

In an embodiment, the invention relates to a miPEP as defined above,said miPEP being labelled.

Without restriction, a labeled peptide may be obtained by fusing saidpeptide to another peptide sequence, to a fluorescent marker (GUS, GFP,FAM, FITC, LacZ . . . ), to a signal sequence (allowing the addressingof the peptide), to a Tag sequence (allowing purification of miPEP) orto a radioactive isotope (I¹²⁵, P³², . . . ).

In an embodiment, the invention relates to a labelled miPEP as definedabove, said labelled miPEP being selected from the group of peptidesconsisting of:

-   -   SEQ ID NO: 1 to SEQ ID NO: 104 and SEQ ID NO: 355,    -   SEQ ID NO: 375 to SEQ ID NO: 386, or    -   SEQ ID NO: 424.

In a particular embodiment, the invention relates to the labelledMtmiPEP171b1 (SEQ ID NO: 59), preferably labelled by a fluorescentmarker such as GUS, GFP, FAM, FITC or LacZ.

In a particular embodiment, the invention relates to the labelledAtmiPEP164a1 (SEQ ID NO: 24), preferably labelled by a fluorescentmarker such as GUS, GFP, FAM, FITC or LacZ.

In a particular embodiment, the invention relates to the labelledAtmiPEP165a (SEQ ID NO: 43), preferably labelled by a fluorescent markersuch as GUS, GFP, FAM, FITC or LacZ.

In a particular embodiment, the invention relates to the labelledAtmiPEP319a1 (SEQ ID NO: 76), preferably labelled by a fluorescentmarker such as GUS, GFP, FAM, FITC or LacZ.

In a particular embodiment, the invention relates to the labelledAtmiPEP319a2 (SEQ ID NO: 77), preferably labelled by a fluorescentmarker such as GUS, GFP, FAM, FITC or LacZ.

In a particular embodiment, the invention relates to the labelledAtmiPEP160b1 (SEQ ID NO: 14), preferably labelled by a fluorescentmarker such as GUS, GFP, FAM, FITC or LacZ.

In a particular embodiment, the invention relates to the labelledMtmiPEP169d (SEQ ID NO: 424), preferably labelled by a fluorescentmarker such as GUS, GFP, FAM, FITC or LacZ.

In a particular embodiment, the invention relates to the labelledMtmiPEP171e (SEQ ID NO: 63), preferably labelled by a fluorescent markersuch as GUS, GFP, FAM, FITC or LacZ.

In a particular embodiment, the invention relates to the labelledDmmiPEP1a (SEQ ID NO: 102), preferably labelled by a fluorescent markersuch as GUS, GFP, FAM, FITC or LacZ.

In a particular embodiment, the invention relates to the labelledDmmiPEP1b (SEQ ID NO: 103), preferably labelled by a fluorescent markersuch as GUS, GFP, FAM, FITC or LacZ.

In a particular embodiment, the invention relates to the labelledDmmiPEP8 (SEQ ID NO: 104), preferably labelled by a fluorescent markersuch as GUS, GFP, FAM, FITC or LacZ.

In a particular embodiment, the invention relates to the labelledHsmiPEP155 (SEQ ID NO: 355), preferably labelled by a fluorescent markersuch as GUS, GFP, FAM, FITC or LacZ.

In another aspect, the invention relates to a nucleic acid moleculeencoding a miPEP as defined above.

In an embodiment, the invention relates to a nucleic acid molecule asdefined above, said molecule being selected from the group of nucleicacids consisting of SEQ ID NO: 105 to SEQ ID NO 208, SEQ ID NO: 387 toSEQ ID NO: 399 and SEQ ID NO: 356 (Table 2).

In an embodiment, the invention relates to a nucleic acid molecule asdefined above, said molecule being selected from the group of nucleicacids consisting of:

-   -   SEQ ID NO: 105 to SEQ ID NO: 208 and SEQ ID NO: 356,    -   SEQ ID NO: 387 to SEQ ID NO: 399, or    -   SEQ ID NO: 425.

In a particular embodiment, the invention relates to MtmiPEP171b1 (SEQID NO: 59) encoded by the nucleotide sequence (SEQ ID NO: 163) containedin the primary transcript of miR171b (SEQ ID NO: 319), said MtmiPEP171b1being capable of modulating the accumulation of said miR171b in aeukaryotic cell.

In a particular embodiment, the invention relates to AtmiPEP164a1 (SEQID NO: 24) encoded by the nucleotide sequence (SEQ ID NO: 128) containedin the primary transcript of miR164a (SEQ ID NO: 297), said AtmiPEP164a1being capable of modulating the accumulation of said miR164a in aeukaryotic cell.

In a particular embodiment, the invention relates to AtmiPEP165a (SEQ IDNO: 43) encoded by the nucleotide sequence (SEQ ID NO: 147) contained inthe primary transcript of miR165a (SEQ ID NO: 305), said miPEP165a beingcapable of modulating the accumulation of said miR165a in a eukaryoticcell.

In a particular embodiment, the invention relates to AtmiPEP319a1 (SEQID NO: 76) encoded by the nucleotide sequence (SEQ ID NO: 180) containedin the primary transcript of miR319a (SEQ ID NO: 331), said AtmiPEP319a1being capable of modulating the accumulation of said miR319a in aeukaryotic cell.

In a particular embodiment, the invention relates to AtmiPEP319a2 (SEQID NO: 77) encoded by the nucleotide sequence (SEQ ID NO: 181) containedin the primary transcript of miR319a (SEQ ID NO: 331), said AtmiPEP319a2being capable of modulating the accumulation of said miR319a in aeukaryotic cell.

In a particular embodiment, the invention relates to AtmiPEP160b1 (SEQID NO: 14) encoded by the nucleotide sequence (SEQ ID NO: 118) containedin the primary transcript of miR160b (SEQ ID NO: 291), said AtmiPEP160b1being capable of modulating the accumulation of said miR160b in aeukaryotic cell.

In a particular embodiment, the invention relates to MtmiPEP169d (SEQ IDNO: 424) encoded by the nucleotide sequence (SEQ ID NO: 425) containedin the primary transcript of miR169d (SEQ ID NO: 427), said MtmiPEP169dbeing capable of modulating the accumulation of said miR169d in aeukaryotic cell.

In a particular embodiment, the invention relates to MtmiPEP171e (SEQ IDNO: 63) encoded by the nucleotide sequence (SEQ ID NO: 167) contained inthe primary transcript of miR171e (SEQ ID NO: 322), said MtmiPEP171ebeing capable of modulating the accumulation of said miR171e in aeukaryotic cell.

In a particular embodiment, the invention relates to DmmiPEPla (SEQ IDNO: 102) encoded by the nucleotide sequence (SEQ ID NO: 206) containedin the primary transcript of miR1 (SEQ ID NO: 353), said dmmiPEP1a beingcapable of modulating the accumulation of said miR1 in a eukaryoticcell.

In a particular embodiment, the invention relates to DmmiPEP1b (SEQ IDNO: 103) encoded by the nucleotide sequence (SEQ ID NO: 207) containedin the primary transcript of miR1 (SEQ ID NO: 353), said dmmiPEP1b beingcapable of modulating the accumulation of said miR1 in a eukaryoticcell.

In a particular embodiment, the invention relates to dmmiPEP8 (SEQ IDNO: 104) encoded by the nucleotide sequence (SEQ ID NO: 208) containedin the primary transcript of miR8 (SEQ ID NO: 354), said dmmiPEP8 beingcapable of modulating the accumulation of said miR8 in a eukaryoticcell.

In a particular embodiment, the invention relates to HsmiPEP155 (SEQ IDNO: 355) encoded by the nucleotide sequence (SEQ ID NO: 356) containedin the primary transcript of miR155 (SEQ ID NO: 358), said HsmiPEP155being capable of modulating the accumulation of said miR155 in aeukaryotic cell.

In another aspect, the invention relates to an isolated peptide, or anisolated and purified peptide, or a synthetic peptide or a recombinantpeptide, comprising or consisting of a sequence identical to that of amiPEP, said miPEP in particular being present naturally in a plant, orin an animal, such as humans.

In another aspect, the invention relates to a vector comprising at leastone nucleic acid molecule as defined above.

In another aspect, the invention also relates to the use of at least:

-   -   a miPEP as defined above,    -   a nucleic acid encoding said miPEP, or    -   a vector containing said nucleic acid,

for modulating the expression of at least one gene in a specifiedeukaryotic cell,

said specified eukaryotic cell being capable of expressing a microRNA,the primary transcript of which contains at least one nucleotidesequence encoding said at least one miPEP and the accumulation of whichis modulated by said at least one miPEP,

the expression of said at least one gene being regulated by saidmicroRNA.

In another aspect, the invention also relates to the use of at least:

-   -   a miPEP of 4 to 100 amino acids, preferably of 4 to 40 amino        acids, encoded by a nucleotide sequence contained in the primary        transcript of a microRNA, said miPEP being capable of modulating        the accumulation of said microRNA in a eukaryotic cell,    -   a nucleic acid encoding said miPEP, or    -   a vector containing said nucleic acid,

for modulating the expression of at least one gene in a specifiedeukaryotic cell, said specified eukaryotic cell being capable ofexpressing a microRNA, the primary transcript of which contains at leastone nucleotide sequence encoding said at least one miPEP and theaccumulation of which is modulated by said at least one miPEP,

the expression of said at least one gene being regulated by saidmicroRNA.

In another aspect, the invention also relates to the use of at least:

-   -   a miPEP of 4 to 100 amino acids, or of 3 amino acids, encoded by        a nucleotide sequence contained in the primary transcript of a        microRNA, said miPEP being capable of modulating the        accumulation of said microRNA in a eukaryotic cell,    -   a nucleic acid encoding said miPEP, or    -   a vector containing said nucleic acid,

for modulating the expression of at least one gene in a specifiedeukaryotic cell, said specified eukaryotic cell being capable ofexpressing a microRNA, the primary transcript of which contains at leastone nucleotide sequence encoding said at least one miPEP and theaccumulation of which is modulated by said at least one miPEP,

the expression of said at least one gene being regulated by saidmicroRNA.

In another aspect, the invention also relates to the use of at least:

-   -   a miPEP of 3 amino acids, encoded by a nucleotide sequence        contained in the primary transcript of a microRNA, said miPEP        being capable of modulating the accumulation of said microRNA in        a eukaryotic cell,    -   a nucleic acid encoding said miPEP, or    -   a vector containing said nucleic acid,

for modulating the expression of at least one gene in a specifiedeukaryotic cell,

said specified eukaryotic cell being capable of expressing a microRNA,the primary transcript of which contains at least one nucleotidesequence encoding said at least one miPEP and the accumulation of whichis modulated by said at least one miPEP,

the expression of said at least one gene being regulated by saidmicroRNA.

The invention is based on the surprising observation made by theinventors that it is possible to modulate the expression of one or moretarget genes of one and the same microRNA by modulating the accumulationof said microRNA using a miPEP.

In an embodiment, the invention relates to the use as defined above inwhich said specified eukaryotic cell is a plant cell.

In an embodiment, the invention relates to the use as defined above inwhich said specified eukaryotic cell is a plant cell of a crucifer suchas Arabidopsis thaliana, of a leguminous plant such as Glycine max(soya), Medicago truncatula and Medicago sativa (alfalfa) or of a plantof the Solanaceae family such as Nicotiana benthamiana (tobacco),Solanum tuberosum (potato), Solanum lycopersicum (tomato) or Solanummelongena (aubergine).

In an embodiment, the invention relates to the use as defined above inwhich said specified eukaryotic cell is an animal cell, in particularhuman.

In an embodiment, the invention relates to the use as defined above inwhich said specified eukaryotic cell is an animal cell, in particularhuman, said miPEP not being used for surgical or therapeutic treatmentof the human body or animal body, nor for modifying the genetic identityof a human being.

In an embodiment, the invention relates to the use as defined above inwhich said specified eukaryotic cell is an animal cell, said miPEP beingused for surgical or therapeutic treatment of the human body or animalbody.

In an embodiment, the invention relates to the use as defined above inwhich said microRNA and said gene are of endogenous origin in saidspecified eukaryotic cell.

In an embodiment, the invention relates to the use as defined above inwhich said microRNA and said gene are of exogenous origin in saidspecified eukaryotic cell, said specified eukaryotic cell containing atleast one vector allowing the expression of said microRNA and of saidgene.

In the invention, the expressions “of endogenous origin” and “ofexogenous origin” are used for distinguishing said microRNAs and/or thegenes of different species, in view of the conservation of the sequencesbetween species.

Thus, the term “of endogenous origin” indicates that the microRNA and/orgene may be present naturally in the cell in question. Other copies ofthe microRNA and/or of the gene of endogenous origin may nevertheless beadded artificially to the cell in question, for example by cloning.

Conversely, the term “of exogenous origin” indicates that the microRNAand/or the gene are never present naturally in the cell in question. Itis a microRNA and/or a gene identified in another cellular type or in anorganism of another species; this microRNA and/or this gene aretherefore necessarily introduced artificially into the cell in question.

In the invention, a genetically transformed cell may therefore contain 2groups of microRNAs and/or of genes potentially similar in terms ofsequence, one of endogenous origin and the other of exogenous origin.

In an embodiment, the invention relates to the use as defined above inwhich the primary transcript of the miRNA and said gene are of exogenousorigin in said specified eukaryotic cell, said specified eukaryotic cellcontaining at least one vector allowing the expression of the primarytranscript of the microRNA.

In an embodiment, the invention relates to the use as defined above inwhich the primary transcript of the miRNA is encoded by a vectorintroduced into the cell artificially.

In an embodiment, the invention relates to the use as defined above inwhich said miPEP is selected from the group of peptides consisting ofSEQ ID NO: 1 to SEQ ID NO: 104, SEQ ID NO: 375 to SEQ ID NO: 386 and SEQID NO: 355 (Table 1).

In an embodiment, the invention relates to the use as defined above inwhich said miPEP is selected from the group of peptides consisting of:

-   -   SEQ ID NO: 1 to SEQ ID NO: 104 and SEQ ID NO: 355,    -   SEQ ID NO: 375 to SEQ ID NO: 386, or    -   SEQ ID NO: 424.

In an embodiment, the invention relates to the use as defined above inwhich said miPEP is selected from MtmiPEP171b1 (SEQ ID NO: 59),AtmiPEP164a1 (SEQ ID NO: 24), AtmiPEP165a (SEQ ID NO: 43), AtmiPEP319a1(SEQ ID NO: 76) and AtmiPEP319a2 (SEQ ID NO: 77).

In an embodiment, the invention relates to the use as defined above inwhich said miPEP is selected from MtmiPEP171b1 (SEQ ID NO: 59),AtmiPEP164a1 (SEQ ID NO: 24), AtmiPEP165a (SEQ ID NO: 43), AtmiPEP319a1(SEQ ID NO: 76), AtmiPEP319a2 (SEQ ID NO: 77), AtmiPEP160b1 (SEQ ID NO:14), MtmiPEP171e (SEQ ID NO: 63) and MtmiPEP169d (SEQ ID NO: 424),In anembodiment, the invention relates to the use as defined above in whichsaid miPEP is selected from DmmiPEPla (SEQ ID NO: 102),

DmmiPEPlb (SEQ ID NO: 103) and DmmiPEP8 (SEQ ID NO: 104).

In an embodiment, the invention relates to the use as defined above inwhich said miPEP is HsmiPEP155a (SEQ ID NO: 355).

In an embodiment, the invention relates to the use as defined above inwhich said nucleic acid is selected from the group of nucleic acidsconsisting of SEQ ID NO: 105 to SEQ ID NO: 208 and SEQ ID NO: 356 (Table2).

In an embodiment, the invention relates to the use as defined above inwhich said nucleic acid is selected from the group of nucleic acidsconsisting of:

-   -   SEQ ID NO: 105 to SEQ ID NO: 208 and SEQ ID NO: 356,    -   SEQ ID NO: 387 to SEQ ID NO: 399, or    -   SEQ ID NO: 425.

In an embodiment, the invention relates to the use as defined above inwhich said nucleic acid is selected from miORF171b (SEQ ID NO: 163),miORF164a1 (SEQ ID NO: 128), miORF165a (SEQ ID NO: 147), miORF319a1 (SEQID NO: 180) and miORF319a2 (SEQ ID NO: 181).

In an embodiment, the invention relates to the use as defined above inwhich said nucleic acid is selected from miORF171b (SEQ ID NO: 163),miORF164a1 (SEQ ID NO: 128), miORF165a (SEQ ID NO: 147), miORF319a1 (SEQID NO: 180), miORF319a2 (SEQ ID NO: 181), miORF160b1 (SEQ ID NO: 118),miORF169d (SEQ ID NO: 425) and miORF171e (SEQ ID NO: 167).

In an embodiment, the invention relates to the use as defined above inwhich said nucleic acid is selected from miORF1a (SEQ ID NO: 206),miORFlb (SEQ ID NO: 207) and miORF8 (SEQ ID NO: 208).

In an embodiment, the invention relates to the use as defined above inwhich said nucleic acid is selected from miORF155 (SEQ ID NO: 356).

In an embodiment, the invention relates to the use as defined above inwhich said microRNA is selected from the group of nucleic acidsconsisting of SEQ ID NO: 282 to SEQ ID NO: 354 and SEQ ID NO: 358.

In an embodiment, the invention relates to the use as defined above inwhich said microRNA is selected from the group of nucleic acidsconsisting of:

-   -   SEQ ID NO: 282 to SEQ ID NO: 354 and SEQ ID NO: 358,    -   SEQ ID NO: 412 to SEQ ID NO: 423, or    -   SEQ ID NO: 427.

In an embodiment, the invention relates to the use as defined above inwhich said microRNA is selected from miR171b (SEQ ID NO: 319), miR165a(SEQ ID NO: 305) and miR319a (SEQ ID NO: 331).

In an embodiment, the invention relates to the use as defined above inwhich said microRNA is selected from miR171b (SEQ ID NO: 319), miR164a(SEQ ID NO: 297), miR165a (SEQ ID NO: 305), miR319a (SEQ ID NO: 331),miR160b (SEQ ID NO: 291), miR171e (SEQ ID NO: 322) and miR169d (SEQ IDNO: 427).

In an embodiment, the invention relates to the use as defined above inwhich said microRNA is selected from miRla (SEQ ID NO: 353) and miR8(SEQ ID NO: 354).

In an embodiment, the invention relates to the use as defined above inwhich said microRNA is selected from miR155 (SEQ ID NO: 358).

In another aspect, the invention relates in particular to a process formodulating the expression of a gene regulated by a microRNA in aeukaryotic cell,

comprising carrying out a step of accumulation of a miPEP in saideukaryotic cell,

-   -   said miPEP having:        -   a size from 3 to 100 amino acids, preferably 4 to 20 amino            acids, and        -   a peptide sequence identical to that encoded by a nucleotide            sequence contained in the primary transcript of a microRNA            regulating the expression of said gene, and        -   being capable of modulating the accumulation of said            microRNA,

in which the accumulation of said miPEP in said eukaryotic cell inducesa modulation of the expression of said gene relative to the expressionof said gene without accumulation of said miPEP.

In another aspect, the invention relates in particular to a process formodulating the expression of a gene regulated by a microRNA in aeukaryotic cell,

comprising carrying out a step of accumulation of a miPEP in saideukaryotic cell,

-   -   said miPEP having:        -   a size from 4 to 100 amino acids, or of 3 amino acids, and        -   a peptide sequence identical to that encoded by a nucleotide            sequence contained in the primary transcript of a microRNA            regulating the expression of said gene, and        -   being capable of modulating the accumulation of said            microRNA,

in which the accumulation of said miPEP in said eukaryotic cell inducesa modulation of the expression of said gene relative to the expressionof said gene without accumulation of said miPEP.

In another aspect, the invention relates in particular to a process formodulating the expression of a gene regulated by a microRNA in aeukaryotic cell,

comprising carrying out a step of accumulation of a miPEP in saideukaryotic cell,

-   -   said miPEP having:        -   a size of 3 amino acids, and        -   a peptide sequence identical to that encoded by a nucleotide            sequence contained in the primary transcript of a microRNA            regulating the expression of said gene, and        -   being capable of modulating the accumulation of said            microRNA,

in which the accumulation of said miPEP in said eukaryotic cell inducesa modulation of the expression of said gene relative to the expressionof said gene without accumulation of said miPEP.

In particular, the invention relates to a process for modulating theexpression of a gene regulated by a microRNA in a eukaryotic cell,

comprising carrying out a step of accumulation of a miPEP in saideukaryotic cell,

-   -   said miPEP having:        -   a size from 4 to 100 amino acids, preferably 4 to 20 amino            acids, and        -   a peptide sequence identical to that encoded by a nucleotide            sequence contained in the primary transcript of a microRNA            regulating the expression of said gene, and        -   being capable of modulating the accumulation of said            microRNA,

in which the accumulation of said miPEP in said eukaryotic cell inducesa modulation of the expression of said gene relative to the expressionof said gene without accumulation of said miPEP.

In an embodiment, the invention relates to a process for modulating theexpression of a gene as defined above, in which the accumulation of saidmiPEP in the cell results from:

-   -   introduction of a nucleic acid encoding said miPEP into the        cell, or    -   introduction of said miPEP into the cell.

In an embodiment, the invention relates to a process for modulating theexpression of a gene as defined above in which said eukaryotic cell is aplant cell.

In an embodiment, the invention relates to a process for modulating theexpression of a gene as defined above in which said eukaryotic cell isan animal cell and in particular a human cell.

In an embodiment, the invention relates to a process for modulating theexpression of a gene as defined above in which said eukaryotic cell isan animal cell and in particular a human cell, said process not beingused for surgical or therapeutic treatment of the human body or animalbody, nor for modifying the genetic identity of a human being.

In an embodiment, the invention relates to a process for modulating theexpression of a gene as defined above in which said microRNA and saidgene are of endogenous origin in said eukaryotic cell.

In an embodiment, the invention relates to a process for modulating theexpression of a gene as defined above in which said microRNA and saidgene are of exogenous origin in said eukaryotic cell, said eukaryoticcell containing at least one vector allowing the expression of saidmicroRNA and of said gene.

In an embodiment, the invention relates to a process for modulating theexpression of a gene as defined above in which said miPEP is selectedfrom the group of peptides consisting of SEQ ID NO: 1 to SEQ ID NO: 104,SEQ ID NO: 375 to SEQ ID NO: 386 and SEQ ID NO: 355.

In an embodiment, the invention relates to a process for modulating theexpression of a gene as defined above in which said miPEP is selectedfrom the group of peptides consisting of:

-   -   SEQ ID NO: 1 to SEQ ID NO: 104 and SEQ ID NO: 355,    -   SEQ ID NO: 375 to SEQ ID NO: 386, or    -   SEQ ID NO: 424.

In a particular embodiment, the invention relates to a process formodulating the expression of a gene regulated by miR171b (SEQ ID NO:319) in a eukaryotic cell, comprising carrying out a step ofaccumulation of MtmiPEP171b1 (SEQ ID NO : 59) in said eukaryotic cell,

in which the accumulation of said MtmiPEP171b1 in said eukaryotic cellinduces a modulation of the expression of said gene relative to theexpression of said gene without accumulation of said MtmiPEP171b1.

In a particular embodiment, the invention relates to a process formodulating the expression of a gene regulated by miR171b (SEQ ID NO:319) in a eukaryotic cell, comprising carrying out a step ofaccumulation of MtmiPEP171b1 (SEQ ID NO: 59) in said eukaryotic cell,

in which the accumulation of said MtmiPEP171b1 in said eukaryotic cellinduces a modulation of the expression of said gene relative to theexpression of said gene without accumulation of said MtmiPEP171b1,

in which said gene is selected from the genes HAM1 (accession No.MtGI9-TC114268) and HAM2 (accession No. MtGI9-TC120850) (accessionnumbers according to the database Medicago truncatula Gene ExpressionAtlas “MtGEA”).

In a particular embodiment, the invention relates to a process formodulating the expression of a gene regulated by miR164a (SEQ ID NO:297) in a eukaryotic cell,

comprising carrying out a step of accumulation of AtmiPEP165a1 (SEQ IDNO: 24) in said eukaryotic cell,

in which the accumulation of said AtmiPEP164a1 in said eukaryotic cellinduces a modulation of the expression of said gene relative to theexpression of said gene without accumulation of said AtmiPEP164a1.

In a particular embodiment, the invention relates to a process formodulating the expression of a gene regulated by miR165a (SEQ ID NO:305) in a eukaryotic cell,

comprising carrying out a step of accumulation of AtmiPEP165a (SEQ IDNO: 43) in said eukaryotic cell,

in which the accumulation of said AtmiPEP165a in said eukaryotic cellinduces a modulation of the expression of said gene relative to theexpression of said gene without accumulation of said AtmiPEP165a.

In a particular embodiment, the invention relates to a process formodulating the expression of a gene regulated by miR319a (SEQ ID NO:331) in a eukaryotic cell, comprising carrying out a step ofaccumulation of AtmiPEP319a1 (SEQ ID NO: 76) in said eukaryotic cell,

in which the accumulation of said AtmiPEP319a1 in said eukaryotic cellinduces a modulation of the expression of said gene relative to theexpression of said gene without accumulation of said AtmiPEP319a1.

In a particular embodiment, the invention relates to a process formodulating the expression of a gene regulated by miR319a (SEQ ID NO:331) in a eukaryotic cell,

comprising carrying out a step of accumulation of AtmiPEP319a2 (SEQ IDNO: 77) in said eukaryotic cell,

in which the accumulation of said AtmiPEP319a2 in said eukaryotic cellinduces a modulation of the expression of said gene relative to theexpression of said gene without accumulation of said AtmiPEP319a2.

In a particular embodiment, the invention relates to a process formodulating the expression of a gene regulated by miR160b (SEQ ID NO:291) in a eukaryotic cell,

comprising carrying out a step of accumulation of AtmiPEP160b1 (SEQ IDNO: 14) in said eukaryotic cell,

in which the accumulation of said AtmiPEP160b1 in said eukaryotic cellinduces a modulation of the expression of said gene relative to theexpression of said gene without accumulation of said AtmiPEP160b1.

In a particular embodiment, the invention relates to a process formodulating the expression of a gene regulated by miR169d (SEQ ID NO:427) in a eukaryotic cell,

comprising carrying out a step of accumulation of MtmiPEP169d (SEQ IDNO: 424) in said eukaryotic cell,

in which the accumulation of said MtmiPEP169d in said eukaryotic cellinduces a modulation of the expression of said gene relative to theexpression of said gene without accumulation of said MtmiPEP169d.

In a particular embodiment, the invention relates to a process formodulating the expression of a gene regulated by miR171e (SEQ ID NO:322) in a eukaryotic cell,

comprising carrying out a step of accumulation of MtmiPEP171e (SEQ IDNO: 63) in said eukaryotic cell,

in which the accumulation of said MtmiPEP171e in said eukaryotic cellinduces a modulation of the expression of said gene relative to theexpression of said gene without accumulation of said MtmiPEP171e.

In a particular embodiment, the invention relates to a process formodulating the expression of a gene regulated by miR1 (SEQ ID NO: 353)in a eukaryotic cell,

comprising carrying out a step of accumulation of DmmiPEP1a (SEQ ID NO:102) in said eukaryotic cell,

in which the accumulation of said DmmiPEPla in said eukaryotic cellinduces a modulation of the expression of said gene relative to theexpression of said gene without accumulation of said DmmiPEP1a.

In a particular embodiment, the invention relates to a process formodulating the expression of a gene regulated by miRl (SEQ ID NO: 353)in a eukaryotic cell,

comprising carrying out a step of accumulation of DmmiPEP1b (SEQ ID NO:103) in said eukaryotic cell,

in which the accumulation of said DmmiPEP1b in said eukaryotic cellinduces a modulation of the expression of said gene relative to theexpression of said gene without accumulation of said DmmiPEP1b.

In a particular embodiment, the invention relates to a process formodulating the expression of a gene regulated by miR8 (SEQ ID NO: 354)in a eukaryotic cell,

comprising carrying out a step of accumulation of DmmiPEP8 (SEQ ID NO:104) in said eukaryotic cell,

in which the accumulation of said DmmiPEP8 in said eukaryotic cellinduces a modulation of the expression of said gene relative to theexpression of said gene without accumulation of said DmmiPEP8.

In a particular embodiment, the invention relates to a process formodulating the expression of a gene regulated by miR155 (SEQ ID NO: 358)in a eukaryotic cell,

comprising carrying out a step of accumulation of hsmiPEP155 (SEQ ID NO:355) in said eukaryotic cell,

in which the accumulation of said hsmiPEP155 in said eukaryotic cellinduces a modulation of the expression of said gene relative to theexpression of said gene without accumulation of said hsmiPEP155.

In another aspect, the invention relates to a modified eukaryotic cellcontaining a peptide identical to a miPEP as defined above, said peptidebeing present in said eukaryotic cell independently of transcription ofthe primary transcript of the microRNA bearing the nucleotide sequenceencoding said miPEP.

In the invention, by the term “modified eukaryotic cell” is meant thatsaid eukaryotic cell contains a miPEP introduced into the cellartificially, whether as a peptide, or via a vector encoding said miPEP.

In addition to a miPEP introduced artificially in the cell, a modifiedeukaryotic cell according to the invention also contains at least onenucleic acid corresponding to the miORF encoding said miPEP.

In addition to a miPEP introduced artificially in the cell and of themiORF, a modified eukaryotic cell according to the invention alsocontains at least one miR, the primary transcript of which contains saidmiORF.

In an embodiment, the invention relates to a modified eukaryotic cell asdefined above, in which said microRNA is of endogenous origin.

In another embodiment, the invention relates to a modified eukaryoticcell as defined above in which said microRNA is of exogenous origin,said modified eukaryotic cell containing a vector allowing theexpression of said microRNA.

In an embodiment, the invention relates to a modified eukaryotic cell asdefined above, said cell being a plant cell.

In an embodiment, the invention relates to a modified eukaryotic cell asdefined above, in which said plant cell is a cell of Medicago truncatulaor of Arabidopsis thaliana, and said peptide is selected from the groupof peptides consisting of:

-   -   SEQ ID NO: 1 to SEQ ID NO: 101,    -   SEQ ID NO: 375 to SEQ ID NO: 386, or    -   SEQ ID NO: 424.

In an embodiment, the invention relates to a modified eukaryotic cell asdefined above, in which said plant cell is a cell of Medicago truncatulaor of Arabidopsis thaliana, and said peptide is selected from the groupconsisting of: SEQ ID NO: 43, SEQ ID NO: 59 and SEQ ID NO: 77.

In an embodiment, the invention relates to a modified eukaryotic cell asdefined above, in which said plant cell is a cell of Medicago truncatulaor of Arabidopsis thaliana, and said peptide is selected from the groupconsisting of: SEQ ID NO: 59, SEQ ID NO: 24, SEQ ID NO: 43, SEQ ID NO:76, SEQ ID NO: 77, SEQ ID NO: 14, SEQ ID NO: 424 and SEQ ID NO: 63.

In an embodiment, the invention relates to a modified eukaryotic cell asdefined above, said cell being an animal cell.

In an embodiment, the invention relates to a modified eukaryotic cell asdefined above, in which said animal cell is a Drosophila cell and saidpeptide is selected from the group of peptides consisting of SEQ ID NO:102, SEQ ID NO: 103 and SEQ ID NO: 104.

In an embodiment, the invention relates to a modified eukaryotic cell asdefined above, in which said animal cell is a human cell and saidpeptide consists of SEQ ID NO: 355.

In another aspect, the invention relates to a plant comprising at leastone modified eukaryotic cell as defined above.

In another aspect, the invention also relates to a non-human animalorganism comprising at least one modified eukaryotic cell as definedabove.

In another aspect, the invention relates to a composition comprising atleast:

-   -   a miPEP as defined above,    -   a nucleic acid encoding said miPEP, or    -   a vector containing said nucleic acid.

In another aspect, the invention relates to a composition comprising atleast one miPEP selected from the group consisting of:

-   -   SEQ ID NO: 1 to SEQ ID NO: 101,    -   SEQ ID NO: 375 to SEQ ID NO: 386, or    -   SEQ ID NO: 424.

In another aspect, the invention relates to a pesticide compositioncomprising at least:

-   -   a miPEP as defined above,    -   a nucleic acid encoding said miPEP, or    -   a vector containing said nucleic acid.

In another aspect, the invention relates to a phytopharmaceuticalcomposition comprising at leaste:

-   -   a miPEP as defined above,    -   a nucleic acid encoding said miPEP, or    -   a vector containing said nucleic acid.

In another aspect, the invention relates to an elicitor compositioncomprising at least:

-   -   a miPEP as defined above,    -   a nucleic acid encoding said miPEP, or    -   a vector containing said nucleic acid.

“Elicitor composition” denotes a composition capable of endowing theplant with better capacity for symbiosis or better resistance todifferent stresses whether they are of the nature of thermal stress,water stress or chemical stress.

For this purpose, the invention also relates to compositions acting onthe growth (inhibition of growth or conversely growth promotion) and thephysiology (better capacity for mycorrhization, nodule formation, bettertolerance of different stresses) of the plant.

In particular, the invention relates to compositions for promoting plantgrowth.

In another aspect, the invention relates to a herbicide compositioncomprising at least:

-   -   a miPEP as defined above,    -   a nucleic acid encoding said miPEP, or    -   a vector containing said nucleic acid.

In another aspect, the invention relates to an insecticide compositioncomprising at least:

-   -   a miPEP as defined above,    -   a nucleic acid encoding said miPEP, or    -   a vector containing said nucleic acid.

In another aspect, the invention relates to a composition, in particulara phytosanitary composition, comprising miPEP164a as active ingredient,said miPEP164a preferably consisting of SEQ ID NO: 24.

In another aspect, the invention relates to a composition, in particulara phytosanitary composition, comprising miPEP319a as active ingredient,said miPEP319a preferably consisting of SEQ ID NO: 76.

In another aspect, the invention relates to a composition, in particulara phytosanitary composition, comprising miPEP171b as active ingredient,said miPEP171b preferably consisting of SEQ ID NO: 59.

In another aspect, the invention relates to a composition, in particulara phytosanitary composition, comprising miPEP165a as active ingredient,said miPEP165a preferably consisting of SEQ ID NO: 43.

In another aspect, the invention relates to a composition, in particulara phytosanitary composition, comprising miPEP319a2 as active ingredient,said miPEP319a2 preferably consisting of SEQ ID NO: 77.

In another aspect, the invention relates to a composition, in particulara phytosanitary composition, comprising miPEP160b1 as active ingredient,said miPEP160b1 preferably consisting of SEQ ID NO: 14.

In another aspect, the invention relates to a composition, in particulara phytosanitary composition, comprising miPEP169d as active ingredient,said miPEP169d preferably consisting of SEQ ID NO: 424.

In another aspect, the invention relates to a composition, in particulara phytosanitary composition, comprising miPEP171e as active ingredient,said miPEP171e preferably consisting of SEQ ID NO: 63.The solubilityproperties of the miPEPs are in particular determined by their aminoacid composition. The hydrophilic miPEPs can be dissolved and packagedin aqueous solutions, such as water. The hydrophobic miPEPs can bedissolved and packaged in solvents, such as organic solvents.

For treatment of plants with the miPEPS, the organic solvents aresolvents that are non-toxic to the plants in small quantities, i.e. theydo not have any harmful effect on the development of the plant.Non-limitatively, the organic solvents may be selected from acetonitrileand acetic acid.

The miPEPs can also be dissolved and packaged in mixtures of organicsolvents, such as for example a mixture of acetonitrile and acetic acid.In particular, the miPEPs may be dissolved in a solution comprising 50%acetonitrile, 10% acetic acid and 40% water (volume/volume/volume).

Preferably, miPEPs 164a and 165a are dissolved in water, and miPEPs 171band 319a are dissolved in a solution comprising 50% acetonitrile, 10%acetic acid and 40% water (volume/volume/volume).

Non-limitatively, the compositions, the pesticide compositions, thephytopharmaceutical compositions, the herbicide compositions and theinsecticide compositions as defined above may comprise 10⁻⁹ M to 10⁻⁴ Mof miPEP, in particular 10⁻⁹ M, 10⁻⁸ M, 10⁻⁷ M, 10⁻⁶ M, 10⁻⁵M or 10⁻⁴ Mof miPEP.

Compositions of higher or lower concentration may also be provideddepending on the applications envisaged. For example, compositionscomprising 10⁻¹ M to 10⁻³ M of miPEP, in particular 10⁻¹ M, 10⁻² M or10⁻³ M of miPEP, may be envisaged in the case where the miPEP has to beadministered to the plant by spreading.

In another aspect, the invention relates to the use of a composition asdefined above, as a herbicide for eradicating plants or slowing theirgrowth, preferably as a herbicide specific to a species or to a genus ofplants.

In another aspect, the invention relates to the use of a composition asdefined above, as a phytopharmaceutical agent,

-   -   for promoting the growth and/or development of plants,

in particular for modulating the physiological parameters of a plant, inparticular the biomass, foliar surface area, flowering, fruit size,production and/or selection of plant seeds, in particular forcontrolling the parthenocarpy or the monoecism of a plant, or formodifying the physiological parameters of plant seeds, in particulargermination, establishment of the root system and resistance to waterstress,

-   -   or for preventing or treating plant diseases,

in particular for promoting resistance to infectious diseases.

In another aspect, the invention relates to the use of a composition asdefined above, for modulating the physiological parameters of a plant,in particular biomass, foliar surface area, or fruit size.

In an embodiment, the invention relates to the use of a composition asdefined above, for thinning of orchards in order to increase fruit size.

In an embodiment, the invention relates to the use of a composition asdefined above, for production and/or selection of plant seeds, saidcomposition being used for controlling the parthenocarpy or themonoecism of a plant.

In an embodiment, the invention relates to the use of a composition asdefined above, said composition being administered to said plant via theleaves or via the roots.

In an embodiment, the invention relates to the use of a composition asdefined above, for production and/or selection of plant seeds.

In an embodiment, the invention relates to the use of a composition asdefined above, in which said composition is used for modifying thephysiological parameters of said plant seeds, in particularestablishment of the root system, germination and resistance to waterstress.

In an embodiment, the invention relates to the use of a composition asdefined above, in which said composition is applied by coating orfilm-coating of said plant seeds.

In another aspect, the invention relates to the use of a composition asdefined above, as a pesticide, for eradicating organisms that areharmful to plants or that might be classified as such,

in particular as insecticide, arachnicide, molluscicide or rodenticide.

In an embodiment, the invention relates to the use of a composition asdefined above, as insecticide.

In an embodiment, the invention relates to the use of a composition asdefined above, for eradicating insect pests.

In an embodiment, the invention relates to the use of a composition asdefined above, for eradicating animal species classified as harmful orliable to be classified as such, in particular the Muridae, inparticular the rat.

In an embodiment, the invention relates to the use of a composition asdefined above, as pesticide for eradicating organisms harmful to plantsor liable to be classified as such,

in particular as insecticide, arachnicide, molluscicide, or rodenticide,

in particular by application of said composition to a plant or to asupport in contact with the plant.

In another aspect, the invention relates to the use of a composition asdefined above, in which said composition is applied to a plant toprotect it against insect pests.

In another aspect, the invention relates to the use of a peptide forpromoting the growth of a plant, said peptide being introduced into theplant, said peptide having an amino acid sequence comprising orconsisting of a sequence identical to that of a miPEP naturally presentin said plant,

said miPEP naturally present in said plant being a peptide of 4 to 100amino acids the sequence of which is encoded by an open reading framelocated on the primary transcript of a miRNA, said miPEP being capableof modulating the accumulation of said miRNA in said plant, said miRNAregulating the expression of at least one gene involved in thedevelopment of the vegetative or reproductive parts of the plant, inparticular the roots, stem, leaves or flowers.

The inventors have surprisingly found that the use of peptides thesequence of which comprises or consists of a sequence identical to thatof miPEPs encoded on the primary transcripts of miRNAs makes it possibleto promote the growth of the plants.

In the invention, the term “plant” refers generally to the whole or partof a plant irrespective of its stage of development (including the plantin the form of a seed or a young shoot), to one or more organs of theplant (for example the leaves, roots, stem, flowers), to one or morecells of the plant, or to a cluster of cells of the plant.

In the invention, the term “growth” refers to the development of thewhole or part of a plant over time. The growth of the plant may thus bedetermined and quantified by monitoring developmental parametersobservable for certain parts, cells or organs of the plant, such as theleaves, roots, stems or flowers.

Non-limitatively, the parameters for determining and quantifying thegrowth of a plant may in particular be:

-   -   the size, surface area, volume, mass and the number of leaves,    -   the size and number of flowers,    -   the size of the stem (or spike),    -   the length and number of roots,    -   the earliness of germination,    -   the earliness of budding,    -   the earliness of floral induction (or floral transition),    -   or also the number of cells.

In the case of leguminous plants, plant growth may also be linked to therate of nodulation, or also to the size and number of nodules on theroots.

Moreover, in the invention, the expression “promote plant growth”, or“improve plant growth”, indicates:

-   -   either an acceleration of development (such as for example a        larger leaf size for a plant at a given point in time relative        to a reference plant),    -   or an increase in development (such as for example a larger leaf        size for a plant that cannot be attained by a reference plant),    -   or an acceleration and an increase in the development of the        plant.

It is important to note that the use according to the invention has theadvantage of being ecological, in comparison with the chemical methodsused conventionally in horticulture or in agriculture, as the miPEP is apeptide that is present naturally in the plant.

The invention also relates to the use of a miPEP introduced into a plantfor promoting its growth,

said miPEP introduced being a peptide comprising, or consisting of, asequence identical to that of a miPEP naturally present in said plant,

said miPEP naturally present is a peptide of 4 to 100 amino acids, thesequence of which is encoded by an open reading frame located at 5′ onthe primary transcript of a miRNA,

said miPEP being capable of modulating the accumulation of said miRNA insaid plant,

said miRNA regulating the expression of at least one gene involved inthe development of the vegetative or reproductive parts of the plant, inparticular the roots, stem, leaves or flowers,

the sum total of the quantity of said miPEP introduced and that of saidmiPEP naturally present being strictly greater than the quantity of saidmiPEP naturally present.

In the invention, the expression “miPEP introduced” refers to a miPEPintroduced into the plant artificially as opposed to the “miPEP presentnaturally in the plant”. The introduction of a miPEP into the planttherefore involves a technological step, which is not a naturalphenomenon and corresponds neither to crossing, nor to selection.

The miPEP introduced may be either a peptide produced outside of theplant (for example an isolated and/or purified peptide, a syntheticpeptide or a recombinant peptide), or a peptide produced in the plantfollowing the non-natural introduction of a nucleic acid encoding saidmiPEP into said plant.

The plant into which the miPEP has not been introduced has a basalquantity of said miPEP, which corresponds to the quantity of said miPEPnaturally present. The use of a miPEP comprising, or consisting of, asequence identical to that of said miPEP leads to an increase in thetotal quantity of miPEP, which modulates the accumulation of the miRNAthe primary transcript of which contains the sequence encoding saidmiPEP.

Moreover, the miPEP introduced is present in the plant and itsintroduction has no impact on its stability.

In an embodiment, the invention relates to the use as defined above, inwhich said gene, involved in the development of the vegetative orreproductive parts of the plant, is selected from the group consistingof: NAC1 (Accession No. AT1G56010.1), NAC4 (Accession No. AT5G07680.1),NAC5 (Accession No. AT5G61430.1), CUC1 (Accession No. AT3G15170.1), CUC2(Accession No. AT5G53950.1), TCP3 (Accession No. AT1G53230.1) and TCP4(Accession No. AT3G15030.1) (accession numbers according to the databaseThe Arabidopsis Information Resource “TAIR”).

In particular, the invention relates to the use as defined above, inwhich said gene involved in the development of the vegetative orreproductive parts of the plant is selected from the group consistingof: NAC1, NAC4, NAC5, CUC1 and CUC2.

In an embodiment, the invention relates to the use as defined above, inwhich said gene involved in the development of the vegetative orreproductive parts of the plant is selected from the group consistingof: TCP3 and TCP4.

In an embodiment, the invention relates to the use as defined above, inwhich said miRNA is selected from miR164a and mir319a.

In an embodiment, the invention relates to the use as defined above, inwhich said miR is selected from miR164a, miR319a, miR171b, miR165a,miR160b, miR169d and miR171e.

In an embodiment, the invention relates to the use as defined above, inwhich said miPEP is selected from AtmiPEP164al, AtmiPEP319al,AtmiPEP319a2, MtmiPEP171b1, AtmiPEP165a1, AtmiPEP160b1, MtmiPEP169d,MtmiPEP 171e.

In particular, the invention relates to the use as defined above, inwhich said miR164a has a nucleotide sequence consisting of SEQ ID NO:297.

In particular, the invention relates to the use as defined above, inwhich said miR164a has a nucleotide sequence having at least 80%identity, preferably at least 90% identity, with the nucleotide sequenceSEQ ID NO: 297.

In an embodiment, the invention relates to the use as defined above, inwhich said miPEP is AtmiPEP164al, in particular in which saidAtmiPEP164a1 has an amino acid sequence consisting of SEQ ID NO: 24.

In particular, the invention relates to the use as defined above, inwhich said miR319a has a nucleotide sequence consisting of SEQ ID NO:331.

In particular, the invention relates to the use as defined above, inwhich said miR319a has a nucleotide sequence having at least 80%identity, preferably at least 90% identity, with the nucleotide sequenceSEQ ID NO: 331.

In an embodiment, the invention relates to the use as defined above, inwhich said miPEP is AtmiPEP319a1, in particular in which saidAtmiPEP319a1 has an amino acid sequence consisting of SEQ ID NO: 76.

In an embodiment, the invention relates to the use as defined above, inwhich said plant is a crucifer such as Arabidopsis thaliana, aleguminous plant such as Glycine max (soya), Medicago truncatula andMedicago sativa (alfalfa) or a plant of the Solanaceae family such asNicotiana benthamiana (tobacco), Solanum tuberosum (potato), Solanumlycopersicum (tomato) or Solanum melongena (aubergine).

In an embodiment, the invention relates to the use as defined above, inwhich said plant is a crucifer.

In an embodiment, the invention relates to the use as defined above, inwhich said plant is Arabidopsis thaliana.

In an embodiment, the invention relates to the use as defined above, forpromoting the growth of an Arabidopsis thaliana plant, in whichAtmiPEP164a1 is introduced into said Arabidopsis thaliana plant, saidAtmiPEP164a1 also being naturally present in said Arabidopsis thalianaplant,

said AtmiPEP164a1 introduced being a peptide the sequence of whichcomprises or consists of a sequence identical to that of saidAtmiPEP164a1 naturally present, said sequence of AtmiPEP164a1 naturallypresent being encoded by an open reading frame located at 5′ on theprimary transcript of the miR164a, which miR164a controls the expressionof at least one gene involved in the development of the vegetative orreproductive parts of Arabidopsis thaliana,

the sum total of the quantity of said AtmiPEP164a1 introduced and thatof said AtmiPEP164a1 naturally present being strictly greater than thequantity of said AtmiPEP164a1 naturally present in said Arabidopsisthaliana plant.

In an embodiment, the invention relates to the use as defined above, forpromoting the growth of an Arabidopsis thaliana plant, in which theAtmiPEP319a1 is introduced into said Arabidopsis thaliana plant, saidAtmiPEP319a1 also being naturally present in said Arabidopsis thalianaplant,

said AtmiPEP319a1 introduced being a peptide the sequence of whichcomprises or consists of a sequence identical to that of saidAtmiPEP319a1 naturally present, said sequence of the AtmiPEP319a1naturally present being encoded by an open reading frame located at 5′on the primary transcript of the miR319a, which miR319a controls theexpression of at least one gene involved in the development of thevegetative or reproductive parts of Arabidopsis thaliana,

the sum total of the quantity of said AtmiPEP319a1 introduced and thatof said AtmiPEP319a1 naturally present being strictly greater than thequantity of said AtmiPEP319a1 naturally present in said Arabidopsisthaliana plant.

In an embodiment, the invention relates to the use as defined above, inwhich said miPEP is introduced into the plant externally, preferably bywatering, by spraying or by adding a fertilizer, a compost, a culturesubstrate or an inert support.

In an embodiment, the invention relates to the use as defined above, inwhich said miPEP is introduced by watering and by spraying.

In an embodiment, the invention relates to the use as defined above, inwhich said miPEP is introduced by watering and by adding a fertilizer.

In an embodiment, the invention relates to the use as defined above, inwhich said miPEP is introduced by spraying and by adding a fertilizer.

In an embodiment, the invention relates to the use as defined above, inwhich said miPEP is introduced, by watering, by spraying and by adding afertilizer.

The inventors have in fact unexpectedly found that it is possible toapply a composition comprising a miPEP directly to the plant in order tomodulate the accumulation of the corresponding miRNA in the plant, whichindicates that the miPEP is captured by the plant.

In an embodiment, the invention relates to the use as defined above, inwhich the plant is treated with a composition comprising 10⁻⁹ M to 10⁻⁴M of said miPEP, in particular 10⁻⁹ M, 10⁻⁸ M, 10⁻⁷ M, 10⁻⁶ M, 10⁻⁵ M or10⁻⁴ M of said miPEP.

Preferably, the compositions have a concentration from 10⁻⁸ M to 10⁻⁵ Mfor application by watering or by spraying on the plant.

In addition, compositions of higher or lower concentration may beenvisaged for treating the plant with the miPEP. As a non-limitativeexample, compositions of higher concentration comprising 10⁻¹ M to 10⁻³M, in particular 10⁻² M of miPEP, may be used in the case where themiPEP introduced exogenously is administered to the plant by spreading.

In an embodiment, the invention relates to the use as defined above, inwhich said miPEP is introduced into the plant by means of a nucleic acidencoding said miPEP, said nucleic acid being introduced into the plant.P In an embodiment, the invention relates to the use as defined above,in which the size of the stem is increased in the plant into which saidmiPEP has been introduced relative to the size of the stem of anidentical plant of the same age into which no miPEP has been introduced,or relative to the size of the stem of an identical plant of the sameage into which said miPEP has not been introduced.

In an embodiment, the invention relates to the use as defined above, inwhich the number of leaves is increased in the plant into which saidmiPEP has been introduced relative to the number of leaves of anidentical plant of the same age into which no miPEP has been introduced,or relative to the number of leaves of an identical plant of the sameage into which said miPEP has not been introduced.

In an embodiment, the invention relates to the use as defined above, inwhich the size of the leaves is increased in the plant into which saidmiPEP has been introduced relative to the size of the leaves of anidentical plant of the same age into which no miPEP has been introduced,or relative to the size of the leaves of an identical plant of the sameage into which said miPEP has not been introduced.

In an embodiment, the invention relates to the use as defined above, inwhich the number of roots is increased in the plant into which saidmiPEP has been introduced relative to the number of roots of anidentical plant of the same age into which no miPEP has been introduced,or relative to the number of roots of an identical plant of the same ageinto which said miPEP has not been introduced.

In an embodiment, the invention relates to the use as defined above, inwhich the length of the roots is increased in the plant into which saidmiPEP has been introduced relative to the length of the roots of anidentical plant of the same age into which no miPEP has been introduced,or relative to the length of the roots of an identical plant of the sameage into which said miPEP has not been introduced.

In an embodiment, the invention relates to the use as defined above, inwhich the rate of nodulation is increased in the plant into which saidmiPEP has been introduced relative to the rate of nodulation of anidentical plant of the same age into which no miPEP has been introduced,or relative to the rate of nodulation of an identical plant of the sameage into which said miPEP has not been introduced.

In an embodiment, the invention relates to the use as defined above, inwhich the number of nodules is increased in the plant into which saidmiPEP has been introduced relative to the number of nodules of anidentical plant of the same age into which no miPEP has been introduced,or relative to the number of nodules of an identical plant of the sameage into which said miPEP has not been introduced.

The increase in the parameters for determining and quantifying growth inthe plant into which the miPEP has been introduced (such as the size ofthe stem, the number and size of the leaves, the number and length ofthe roots, the rate of nodulation or also the number of nodules on theroots) is preferably demonstrated by comparison with an identical plant(i.e. a plant of the same species and/or variety), of the same age andgrown under the same conditions but into which no miPEP has beenintroduced.

In another aspect, the invention relates to a process for promoting thegrowth of a plant, comprising a step of introducing a miPEP into aplant, said miPEP also being present naturally in said plant,

said miPEP introduced being a peptide of 4 to 100 amino acids thesequence of which comprises or consists of a sequence identical to thatof said miPEP naturally present, said sequence of the miPEP naturallypresent being encoded by an open reading frame located at 5′ on theprimary transcript of a miRNA, said miPEP being capable of modulatingthe accumulation of said miRNA, said miRNA regulating the expression ofat least one gene involved in the development of the vegetative orreproductive parts of the plant, in particular the roots, stem, leavesor flowers,

the sum total of the quantity of said miPEP introduced and that of saidmiPEP naturally present being strictly greater than the quantity of saidmiPEP naturally present.

In an embodiment, the invention relates to a process as defined above,in which said gene involved in the development of the vegetative orreproductive parts of the plant is selected from the group consistingof: NAC1 (Accession No. AT1G56010.1), NAC4 (Accession No. AT5G07680.1),NAC5 (Accession No. AT5G61430.1), CUC1 (Accession No. AT3G15170.1), CUC2(Accession No. AT5G53950.1), TCP3 (Accession No. AT1G53230.1) and TCP4(Accession No. AT3G15030.1).

In particular, the invention relates to a process as defined above, inwhich said gene involved in the development of the vegetative orreproductive parts of the plant is selected from the group consistingof: NAC1, NAC4, NAC5, CUC1 and CUC2.

In an embodiment, the invention relates to a process as defined above,in which said gene involved in the development of the vegetative orreproductive parts of the plant is selected from the group consistingof: TCP3 and TCP4.

In an embodiment, the invention relates to a process as defined above,in which said miR is selected from miR164a, miR319a, miR171b, miR165a,miR160b, miR169d and miR171e.

In an embodiment, the invention relates to a process as defined above,in which said miPEP is selected from AtmiPEP164a1, AtmiPEP319a1,AtmiPEP319a2, MtmiPEP171b1, AtmiPEP165a1, AtmiPEP160b1, MtmiPEP169d,MtmiPEP171e.

In an embodiment, the invention relates to a process as defined above,in which said miRNA is miR164a, in particular in which said miR164a hasa nucleotide sequence consisting of SEQ ID NO: 297.

In an embodiment, the invention relates to a process as defined above,in which said miPEP is AtmiPEP164a1, in particular in which saidAtmiPEP164a1 has an amino acid sequence consisting of SEQ ID NO: 24.

In an embodiment, the invention relates to a process as defined above,in which said miRNA is miR319a, in particular in which said miR319a hasa nucleotide sequence consisting of SEQ ID NO: 331.

In an embodiment, the invention relates to a process as defined above,in which said miPEP is AtmiPEP319a1, in particular in which saidAtmiPEP319a1 has an amino acid sequence consisting of SEQ ID NO: 76.

In an embodiment, the invention relates to a process as defined above,in which said plant is a crucifer such as Arabidopsis thaliana, aleguminous plant such as Glycine max (soya), Medicago truncatula andMedicago sativa (alfalfa) or a plant of the Solanaceae family such asNicotiana benthamiana (tobacco), Solanum tuberosum (potato), Solanumlycopersicum (tomato) or Solanum melongena (aubergine).

In an embodiment, the invention relates to a process as defined above,in which said plant is a crucifer.

In an embodiment, the invention relates to a process as defined above,in which said plant is Arabidopsis thaliana.

In an embodiment, the invention relates to a process as defined above,for promoting the growth of an Arabidopsis thaliana plant, in whichAtmiPEP164a1 is introduced into said Arabidopsis thaliana plant, saidAtmiPEP164a1 also being naturally present in said Arabidopsis thalianaplant,

said AtmiPEP164a1 introduced being a peptide comprising or consisting ofa sequence identical to that of said AtmiPEP164a1 naturally present,where the AtmiPEP164a1 naturally present is a peptide of 4 to 100 aminoacids the sequence of which is encoded by an open reading frame locatedat 5′ on the primary transcript of the miR164a,

said AtmiPEP164a1 being capable of increasing the accumulation of saidmiR164a, where said miR164a regulates the expression of at least onegene involved in the development of the vegetative or reproductive partsof Arabidopsis thaliana,

the sum total of the quantity of said AtmiPEP164a1 introduced and thatof said AtmiPEP164a1 naturally present being strictly greater than thequantity of said AtmiPEP164a1 naturally present.

In an embodiment, the invention relates to a process as defined above,for promoting the growth of an Arabidopsis thaliana plant, in whichAtmiPEP319a1 is introduced into said Arabidopsis thaliana plant, saidAtmiPEP319a1 also being naturally present in said Arabidopsis thalianaplant,

said AtmiPEP319a1 introduced being a peptide comprising or consisting ofa sequence identical to that of said AtmiPEP319a1 naturally present,where the AtmiPEP319a1 naturally present is a peptide of 4 to 100 aminoacids the sequence of which is encoded by an open reading frame locatedat 5′ on the primary transcript of the miR319a,

said AtmiPEP319a1 being capable of increasing the accumulation of saidmiR319a, where miR319a regulates the expression of at least one geneinvolved in the development of the vegetative or reproductive parts ofArabidopsis thaliana,

the sum total of the quantity of said AtmiPEP319a1 introduced and thatof said AtmiPEP319a1 naturally present being strictly greater than thequantity of said AtmiPEP319a1 naturally present.

In an embodiment, the invention relates to a process as defined above,in which said miPEP is introduced into the plant externally, preferablyby watering, by spraying or by adding a fertilizer, a compost, a culturesubstrate or an inert support.

In an embodiment, the invention relates to a process as defined above,in which said miPEP is administered to the plant in the form of acomposition comprising 10⁻⁹ M to 10⁻⁴ M of said miPEP, in particular10⁻⁹, 10⁻⁸, 10⁻⁷, 10⁻⁶, 10⁻⁵ or 10⁻⁴ M of said miPEP.

In an embodiment, the invention relates to a process as defined above,in which said miPEP is introduced into the plant by means of a nucleicacid encoding said miPEP, said nucleic acid being introduced into theplant.

In an embodiment, the invention relates to a process as defined above,in which the size of the stem is increased in the plant into which saidmiPEP has been introduced relative to the size of the stem of anidentical plant of the same age into which no miPEP has been introduced,or relative to the size of the stem of an identical plant of the sameage into which said miPEP has not been introduced.

In an embodiment, the invention relates to a process as defined above,in which the number of leaves is increased in the plant into which saidmiPEP has been introduced relative to the number of leaves of anidentical plant of the same age into which no miPEP has been introduced,or relative to the number of leaves of an identical plant of the sameage into which said miPEP has not been introduced.

In an embodiment, the invention relates to a process as defined above,in which the size of the leaves is increased in the plant into whichsaid miPEP has been introduced relative to the size of the leaves of anidentical plant of the same age into which no miPEP has been introduced,or relative to the size of the leaves of an identical plant of the sameage into which said miPEP has not been introduced.

In an embodiment, the invention relates to a process as defined above,in which the number of roots is increased in the plant into which saidmiPEP has been introduced relative to the number of roots of anidentical plant of the same age into which no miPEP has been introduced,or relative to the number of roots of an identical plant of the same ageinto which said miPEP has not been introduced.

In an embodiment, the invention relates to a process as defined above,in which the length of the roots is increased in the plant into whichsaid miPEP has been introduced relative to the length of the roots of anidentical plant of the same age into which no miPEP has been introduced,or relative to the length of the roots of an identical plant of the sameage into which said miPEP has not been introduced.

In an embodiment, the invention relates to a process as defined above,in which the rate of nodulation is increased in the plant into whichsaid miPEP has been introduced relative to the rate of nodulation of anidentical plant of the same age into which no miPEP has been introduced,or relative to the rate of nodulation of an identical plant of the sameage into which said miPEP has not been introduced.

In an embodiment, the invention relates to a process as defined above,in which the number of nodules is increased in the plant into which saidmiPEP has been introduced relative to the number of nodules of anidentical plant of the same age into which no miPEP has been introduced,or relative to the number of nodules of an identical plant of the sameage into which said miPEP has not been introduced.

In another aspect, the invention relates to a plant into which a miPEPhas been introduced according to the use or the process for promotingthe growth of a plant described above. In another aspect, the inventionrelates to a process for producing a transgenic plant comprising: a) astep of introducing a nucleic acid encoding a miPEP of 4 to 100 aminoacids into a plant, or into at least one cell of said plant, underconditions allowing the expression of said miPEP,

said miPEP also being naturally present in said plant, said miPEPnaturally present being a peptide the sequence of which is encoded by anopen reading frame located at 5′ on the primary transcript of a miR,said miPEP being capable of modulating the accumulation of said miR inthe plant, said miR regulating the expression of at least one geneinvolved in the development of the vegetative or reproductive parts ofthe plant, in particular the roots, stem, leaves or flowers, and

-   b) a step of culturing the plant, or at least one cell of said    plant, obtained in step a) under conditions allowing a transgenic    plant to be obtained.

In another aspect, the invention relates to a process for producing atransgenic plant comprising:

-   a) a step of introducing a nucleic acid encoding a miPEP of 4 to 100    amino acids, or of 3 amino acids, into a plant, or into at least one    cell of said plant, under conditions allowing the expression of said    miPEP,

said miPEP also being naturally present in said plant, said miPEPnaturally present being a peptide the sequence of which is encoded by anopen reading frame located at 5′ on the primary transcript of a miR,said miPEP being capable of modulating the accumulation of said miR inthe plant, said miR regulating the expression of at least one geneinvolved in the development of the vegetative or reproductive parts ofthe plant, in particular the roots, stem, leaves or flowers, and

-   b) a step of culturing the plant, or at least one cell of said    plant, obtained in step a) under conditions allowing a transgenic    plant to be obtained.

In another aspect, the invention relates to a process for producing atransgenic plant comprising:

-   a) a step of introducing a nucleic acid encoding a miPEP of 3 amino    acids, into a plant, or into at least one cell of said plant, under    conditions allowing the expression of said miPEP,

said miPEP also being naturally present in said plant, said miPEPnaturally present being a peptide the sequence of which is encoded by anopen reading frame located at 5′ on the primary transcript of a miR,said miPEP being capable of modulating the accumulation of said miR inthe plant, said miR regulating the expression of at least one geneinvolved in the development of the vegetative or reproductive parts ofthe plant, in particular the roots, stem, leaves or flowers, and

b) a step of culturing the plant, or at least one cell of said plant,obtained in step a) under conditions allowing a transgenic plant to beobtained.

In an embodiment, the invention relates to a process for producing atransgenic plant as defined above, in which said transgenic plantobtained in step b) has improved growth relative to an identical plantin which said nucleic acid has not been introduced.

In an embodiment, the invention relates to a process for producing atransgenic plant as defined above, in which the expression of saif miPEPencoded by the nucleic acid introduced in the plant results in animproved growth relative to an identical plant in which said nucleicacid has not been introduced.

In an embodiment, the invention relates to a process for producing atransgenic plant as defined above, in which step a) is carried out usinga vector containing said nucleic acid, preferably a plasmid.

In an embodiment, the invention relates to a process for producing atransgenic plant as defined above, in which the expression of saidnucleic acid of step a) is placed under the control of a strongpromoter, preferably a constitutive strong promoter such as the 35Spromoter.

In an embodiment, the invention relates to a process for producing atransgenic plant as defined above, in which said gene involved in thedevelopment of the vegetative or reproductive parts of the plant isselected from the group consisting of: NAC1 (Accession No. AT1G56010.1),NAC4 (Accession No. AT5G07680.1), NAC5 (Accession No. AT5G61430.1), CUC1(Accession No. AT3G15170.1), CUC2 (Accession No. AT5G53950.1), TCP3(Accession No. AT1G53230.1) and TCP4 (Accession No. AT3G15030.1).

In an embodiment, the invention relates to a process for producing atransgenic plant as defined above, in which said miPEP has an amino acidsequence comprising or consisting of a sequence selected from the groupconsisting of SEQ ID NO: 1 to SEQ ID NO: 101 and SEQ ID NO: 375 to SEQID NO: 386.

In an embodiment, the invention relates to a process for producing atransgenic plant as defined above, in which said miPEP has an amino acidsequence comprising or consisting of a sequence selected from the groupconsisting of:

-   -   SEQ ID NO: 1 to SEQ ID NO: 101,    -   SEQ ID NO: 375 to SEQ ID NO: 386, or    -   SEQ ID NO: 424.

In an embodiment, the invention relates to a process for producing atransgenic plant as defined above, in which said miRNA is miR164a, inparticular in which said miR164a has a nucleotide sequence consisting ofSEQ ID NO: 297.

In an embodiment, the invention relates to a process for producing atransgenic plant as defined above, in which said miPEP is AtmiPEP164a1,in particular in which said AtmiPEP164a1 has an amino acid sequenceconsisting of SEQ ID NO: 24.

In an embodiment, the invention relates to a process for producing atransgenic plant as defined above, in which said miRNA is miR319a, inparticular in which said miR319a has a nucleotide sequence consisting ofSEQ ID NO: 331.

In an embodiment, the invention relates to a process for producing atransgenic plant as defined above, in which said miPEP is AtmiPEP319a1,in particular in which said AtmiPEP319a1 has an amino acid sequenceconsisting of SEQ ID NO: 76.

In an embodiment, the invention relates to a process for producing atransgenic plant as defined above, in which said nucleic acid introducedin step a) comprises a nucleotide sequence selected from SEQ ID NO: 128and SEQ ID NO: 180.

In an embodiment, the invention relates to a process for producing atransgenic plant as defined above, in which said plant is a crucifersuch as Arabidopsis thaliana, a leguminous plant such as Glycine max(soya), Medicago truncatula and Medicago sativa (alfalfa) or a plant ofthe Solanaceae family such as Nicotiana benthamiana (tobacco), Solanumtuberosum (potato), Solanum lycopersicum (tomato) or Solanum melongena(aubergine).

In an embodiment, the invention relates to a process for producing atransgenic plant as defined above, in which said transgenic plant is acrucifer.

In an embodiment, the invention relates to a process for producing atransgenic plant as defined above, in which said transgenic plant isArabidopsis thaliana.

In an embodiment, the invention relates to a process for producing atransgenic plant as defined above, comprising:

-   a) a step of introducing a nucleic acid containing the nucleotide    sequence SEQ ID NO: 128, encoding AtmiPEP164a1 consisting of the    amino acid sequence SEQ ID NO: 24, into an Arabidopsis thaliana    plant, or into at least one cell of said Arabidopsis thaliana plant,    under conditions allowing the expression of AtmiPEP164a1,

said AtmiPEP164a1 also being naturally present in said Arabidopsisthaliana plant, said miPEP naturally present being a peptide thesequence of which is encoded by an open reading frame located at 5′ onthe primary transcript of miR164a, said AtmiPEP164a1 being capable ofmodulating the accumulation of said miR164, where miR164a controls theexpression of at least one gene involved in the development of thevegetative or reproductive parts of Arabidopsis thaliana, and

-   b) a step of culturing the plant, or at least one cell of said    plant, obtained in step a) under conditions allowing a transgenic    Arabidopsis thaliana plant to be obtained.

In an embodiment, the invention relates to a process for producing atransgenic plant as defined above, comprising:

-   a) a step of introducing a nucleic acid containing the nucleotide    sequence SEQ ID NO: 180, encoding AtmiPEP319a1 consisting of the    amino acid sequence SEQ ID NO: 76, into an Arabidopsis thaliana    plant, or into at least one cell of said Arabidopsis thaliana plant,    under conditions allowing the expression of AtmiPEP319a1,

said AtmiPEP319a1 also being naturally present in said Arabidopsisthaliana plant, said miPEP naturally present being a peptide thesequence of which is encoded by an open reading frame located at 5′ onthe primary transcript of the miR319a, said AtmiPEP319a1 being capableof modulating the accumulation of said miR319, which miR319a regulatesthe expression of at least one gene involved in the development of thevegetative or reproductive parts of Arabidopsis thaliana, and

-   b) a step of culturing the plant, or at least one cell of said    plant, obtained in step a) under conditions allowing a transgenic    Arabidopsis thaliana plant to be obtained.

In an embodiment, the invention relates to a process of production asdefined above, in which said miPEP is introduced into the plant by meansof a nucleic acid encoding said miPEP, said nucleic acid beingintroduced into the plant.

In an embodiment, the invention relates to a process for producing atransgenic plant as defined above, in which the size of the stem isincreased in the plant into which said miPEP has been introducedrelative to the size of the stem of an identical plant of the same ageinto which no miPEP has been introduced, or relative to the size of thestem of an identical plant of the same age into which said miPEP has notbeen introduced.

In an embodiment, the invention relates to a process for producing atransgenic plant as defined above, in which the number of leaves isincreased in the plant into which said miPEP has been introducedrelative to the number of leaves of an identical plant of the same ageinto which no miPEP has been introduced, or relative to the number ofleaves of an identical plant of the same age into which said miPEP hasnot been introduced.

In an embodiment, the invention relates to a process for producing atransgenic plant as defined above, in which the size of the leaves isincreased in the plant into which said miPEP has been introducedrelative to the size of the leaves of an identical plant of the same ageinto which no miPEP has been introduced, or relative to the size of theleaves of an identical plant of the same age into which said miPEP hasnot been introduced.

In an embodiment, the invention relates to a process for producing atransgenic plant as defined above, in which the number of roots isincreased in the plant into which said miPEP has been introducedrelative to the number of roots of an identical plant of the same ageinto which no miPEP has been introduced, or relative to the number ofroots of an identical plant of the same age into which said miPEP hasnot been introduced.

In an embodiment, the invention relates to a process for producing atransgenic plant as defined above, in which the length of the roots isincreased in the plant into which said miPEP has been introducedrelative to the length of the roots of an identical plant of the sameage into which no miPEP has been introduced, or relative to the lengthof the roots of an identical plant of the same age into which said miPEPhas not been introduced.

In an embodiment, the invention relates to a process for producing atransgenic plant as defined above, in which the rate of nodulation isincreased in the plant into which said miPEP has been introducedrelative to the rate of nodulation of an identical plant of the same ageinto which no miPEP has been introduced, or relative to the rate ofnodulation of an identical plant of the same age into which said miPEPhas not been introduced.

In an embodiment, the invention relates to a process for producing atransgenic plant as defined above, in which the number of nodules isincreased in the plant into which said miPEP has been introducedrelative to the number of nodules of an identical plant of the same ageinto which no miPEP has been introduced, or relative to the number ofnodules of an identical plant of the same age into which said miPEP hasnot been introduced.

In an aspect, the invention also relates to a transgenic plant asobtained by the process of production as defined above.

In another aspect, the invention relates to a transgenic plantcontaining a vector encoding a miPEP, said miPEP being also naturallypresent in said plant,

said miPEP naturally present being a peptide of which the sequence isencoded by an open reading frame located in 5′ portion on the primarytranscript of a miR, said miPEP being capable of modulating theaccumulation of said miR in the plant, said miR regulating theexpression of at least one gene,

the expression of said miPEP encoded by the vector introduced in theplant resulting in a modulation of the expression of said gene relativeto an identical plant in which said nucleic acid has not beenintroduced.

In an embodiment, the invention relates to a transgenic plant as definedabove, in which said gene is involved in the development of thevegetative or reproductive parts of the plant, in particular the roots,stem, leaves or flowers, and the expression of said miPEP encoded by thenucleic acid introduced in the plant results in an improved growth ofthe transgenic plant relative to an identical plant in which saidnucleic acid has not been introduced.

In an embodiment, the invention relates to a transgenic plant as definedabove, in which said miPEP is selected from the group consisting of:

-   -   SEQ ID NO: 1 to SEQ ID NO: 101,    -   SEQ ID NO: 375 to SEQ ID NO: 386, or    -   SEQ ID NO: 424.

In another aspect, the invention relates to a transgenic plantcomprising a vector,

said vector containing a nucleic acid enabling the expression of aprimary transcript of a miR,

said primary transcript of a miR containing an open reading frameencoding a miPEP,

said miPEP being capable of modulating the accumulation of said miR insaid plant, said miR regulating the expression of at least one gene insaid plant,

the expression of the miPEP encoded by the vector introduced in theplant resulting in a modulation of the expression of said gene relativeto an identical plant in which said nucleic acid has not beenintroduced.

In an embodiment, the invention relates to a transgenic plant as definedabove in which said nucleic acid is of endogenous origin.

If said nucleic acid is of endogenous origin, at least one copy of saidnucleic acid is already present naturally in the plant, independently ofthe vector introduced in the plant.

In an embodiment, the invention relates to a transgenic plant as definedabove in which said nucleic acid is of exogenous origin.

If said nucleic acid is of exogenous origin, no copy of said nucleicacid is already present naturally in the plant

In an embodiment, the invention relates to a transgenic plant as definedabove in which said gene is involved in the development of thevegetative or reproductive parts of the plant, in particular the roots,stem, leaves or flowers, and the expression of said miPEP encoded by thenucleic acid introduced in the plant results in an improved growth ofthe transgenic plant relative to an identical plant in which saidnucleic acid has not been introduced.

In an embodiment, the invention relates to a transgenic plant as definedabove in which said miPEP is selected from the group consisting of:

-   -   SEQ ID NO: 1 to SEQ ID NO: 101,    -   SEQ ID NO: 375 to SEQ ID NO: 386, or    -   SEQ ID NO: 424.

In another aspect, the invention relates to a composition comprising, incombination, a quantity of seeds of a plant and a quantity of a peptidethe sequence of which comprises or consists of a sequence identical tothat of a miPEP naturally present in said plant.

In an embodiment, the invention relates to a composition as definedabove in which said miPEP is selected from the group consisting of:

-   -   SEQ ID NO: 1 to SEQ ID NO: 101,    -   SEQ ID NO: 375 to SEQ ID NO: 386, or    -   SEQ ID NO: 424.

In an embodiment, the invention relates to a composition comprising, incombination, a quantity of seeds of a plant, in particular A. thaliana,and a quantity of a peptide the sequence of which comprises or consistsof a sequence identical to that of AtmiPEP164a1.

In an embodiment, the invention relates to a composition comprising, incombination, a quantity of seeds of a plant, in particular A. thaliana,and a quantity of a peptide the sequence of which comprises or consistsof a sequence identical to that of AtmiPEP319a1.

In an embodiment, the invention relates to a composition comprising, incombination, a quantity of seeds of a plant, in particular M truncatula,and a quantity of a peptide the sequence of which comprises or consistsof a sequence identical to that of MtmiPEP171b.

In an embodiment, the invention relates to a composition as definedabove, formulated so as to form a coated seed.

Coating may be carried out by the processes used conventionally in theagri-food industry and may be obtained using a material able todisaggregate in a solvent or in the ground, such as a binder or clay.

According to the invention, coating may be used for example forconferring particular properties on a composition of miPEP, or on acomposition of seeds in combination with a miPEP.

In an embodiment, the invention relates to a composition as definedabove, formulated so as to form a coated seed comprising MtmiPEP171b.

In an embodiment, the invention relates to a composition as definedabove, formulated so as to form a coated seed comprising AtmiPEP164a1.

In an embodiment, the invention relates to a composition as defined,above formulated so as to form a coated seed comprising AtmiPEP319a1.

In another aspect, the invention relates to a pharmaceutical compositioncomprising at least:

-   -   a miPEP as defined above    -   a nucleic acid encoding said miPEP, or    -   a vector containing said nucleic acid.

and a pharmaceutically acceptable excipient.In another aspect, theinvention relates to a composition comprising at least:

-   -   a miPEP as defined above    -   a nucleic acid encoding said miPEP, or    -   a vector containing said nucleic acid.

for use as a medicament, in particular for humans or for animals.

The use of the compositions of the invention is applicable in humanmedicine and in veterinary medicine.

In another aspect, the invention relates to a composition comprising atleast:

-   -   a miPEP as defined above    -   a nucleic acid encoding said miPEP, or    -   a vector containing said nucleic acid,

for use in the prevention and/or treatment of a disease involvingderegulation of the expression of a gene of the patient,

the expression of said gene being regulated by a microRNA theaccumulation of which is modulated by said miPEP.

In an embodiment, the invention relates to the composition as definedabove in which said disease is selected from cancer, diabetes, obesity,infectious diseases and neurodegenerative diseases.

In an embodiment, the invention relates to the composition as definedabove in which said disease is selected from: cancer, diabetes, obesity,infectious diseases, neurodegenerative diseases, cardiovascular diseasesand autoimmune diseases.

In an embodiment, the invention relates to the composition as definedabove in which said disease is selected from: cancer, diabetes, obesity,infectious diseases, neurodegenerative diseases, or from cardiovasculardiseases and autoimmune diseases.

In an embodiment, the invention relates to the composition as definedabove in which said disease is selected from cardiovascular diseases andautoimmune diseases.

In another aspect, the invention relates to a composition comprising atleast:

-   -   a miPEP as defined above,    -   a nucleic acid encoding said miPEP, or    -   a vector containing said nucleic acid,

for use in the prevention and/or treatment of an infection of an animalor of a human with a parasitic organism,

said parasitic organism having a gene the expression of which isregulated by a microRNA the accumulation of which is modulated by saidmiPEP.

In another aspect, the invention relates to a miPEP from 4 to 100 aminoacids, or of 3 amino acids, encoded by a nucleotidic sequence containedin the preimary transcript of a microRNA, said miPEP being capable ofmodulating the accumulation of said microRNA in an eukaryotic cell, forits use a medicament.

In another aspect, the invention relates to an antibody specificallyrecognizing a miPEP.

In particular, the invention relates to an antibody specificallyrecognizing AtmiPEP165a.

In particular, the invention relates to an antibody specificallyrecognizing MtmiPEP171b.

In particular, the invention relates to an antibody specificallyrecognizing AtmiPEP164a1.

In particular, the invention relates to an antibody specificallyrecognizing AtmiPEP319a1.

Such an antibody may be obtained by a process known to a person skilledin the art, such as for example by injecting said miPEP into a non-humananimal in order to trigger an immunization reaction and the productionof antibodies by said animal.

In another aspect, the invention relates to a process ofimmunolocalization of a miPEP comprising a step of labelling abiological sample from a plant with an antibody specifically recognizinga miPEP.

In particular, the invention relates to a process of immunolocalizationof AtmiPEP165a using an antibody specifically recognizing AtmiPEP165a.

In particular, the invention relates to a process of immunolocalizationof MtmiPEP171b using an antibody specifically recognizing MtmiPEP171b.

In particular, the invention relates to a process of immunolocalizationof AtmiPEP164a1 using an antibody specifically recognizing AtmiPEP164a1.

In particular, the invention relates to a process of immunolocalizationof MtmiPEP319a1 using an antibody specifically recognizing AtmiPEP319a1.

In another aspect, the invention relates to an antibody specificallyrecognizing a miPEP according to the invention, for its use as amedicament.

In another aspect, the invention relates a pharmaceutical compositioncomprising an antibody specifically recognizing a miPEP according to theinvention and a pharmaceutically acceptable excipient.

In another aspect, the invention relates to a protocol for producing arecombinant peptide, the sequence of which comprises or consists of asequence identical to that of a miPEP as defined above, comprising astep of transforming an organism with an expression vector encoding saidrecombinant peptide.

In an embodiment, said organism is selected from the group comprisingbacteria, yeasts, fungi (other than yeasts), animal cells, plants andanimals.

In an embodiment, said organism is Escherichia coli.

In particular, the invention relates to a protocol for producing arecombinant peptide as defined above, comprising the following steps:

-   -   binding the nucleic acid encoding said recombinant peptide to a        nucleic acid encoding a tag, such as GST,    -   introducing the expression vector containing said nucleic acid        encoding said recombinant peptide into the bacterium E. coli,    -   culturing the bacterium E. coli containing the expression vector        in LB medium preferably up to an OD between 0.2 and 0.4,    -   inducing production of the recombinant peptide with IPTG,        preferably for 4 to 5 hours,    -   centrifuging and lysing the E. coli bacteria,    -   filtering the supernatant,    -   purifying said recombinant peptide on a glutathione sepharose        affinity column,    -   if necessary, cleaving the GST with a protease.

All the sequences of the miPEPs, miORFs, miRNAs and primary transcriptsof miRNAs are presented in Tables 1, 2, 3, 4, 5 and 6.

Table 7 presents an analysis of the polymorphism of the DNA sequence ofthe different regions of pri-miR171b (a haplotype is defined when itdiffers by at least one amino acid from the other haplotypes).

TABLE 1 List of potential miPEPs (miPEPs) miPEP Target genes ofSequence of (pI/size MW) miRNA Organism the miRNA the miPEP SEQ IDAtmiPEP156a1 miR156a Arabidopsis SPL gene family,MFCSIQCVARHLFPLHVREIKKATRAI SEQ ID NO: 1 (10.57/3824) thalianainvolved in KKGKTL AtmiPEP156a2 miR156a Arabidopsis  development of theMRRQTSVPFACKRDKESDKSHKER SEQ ID NO: 2 thaliana stem and floweringAtmiPEP156a3 miR156a Arabidopsis  MVMFFLDLDKNPRFDLLKGLKWNLF SEQ ID NO: 3thaliana SSHISPSLPPSL AtmiPEP156c1 miR156c Arabidopsis  MKDNFPLLLRLSEQ ID NO: 4 (8.5/1359) thaliana AtmiPEP156c2 miR156c Arabidopsis  MSDDSEQ ID NO: 5 thaliana AtmiPEP156e1 miR156e Arabidopsis  MIYINKYGSISAVEDDSEQ ID NO: 6 (4.03/1818) thaliana AtmiPEP156f1 miR156f Arabidopsis  MSQRSEQ ID NO: 7 (9.5/520) thaliana AlmiPEP159a miR159a Arabidopsis MYB gene family MTCPLLSLSFLLSKYI SEQ ID NO: 8 lyrata involved inAtmiPEP159a1 miR159a Arabidopsis  germination and MTWPLLSLSFLLSKYVSEQ ID NO: 9 (8.34/1898) thaliana flowering CrmiPEP159a miR159aCapsella  MTCTLSALSLSLNMFRVN SEQ ID NO: 10 rubella AtmiPEP159b1 miR159bArabidopsis  MFYLS SEQ ID NO: 11 (5.27/659) thaliana AtmiPEP159b2miR159b Arabidopsis  MVNTSSFFISSFILPLVLSESNCLLFRTI SEQ ID NO: 12thaliana YKFSMVLY AtmiPEP160a1 miR160a Arabidopsis  ARF gene family,MFCLLIPIFSFVFSPNRHLRLQEQ SEQ ID NO: 13 (8.02/2936) thaliana involved inAtmiPEP160b1 miR160b Arabidopsis  germination MFSPQ SEQ ID NO: 14(5.28/608) thaliana development and AtmiPEP160b2 miR160b Arabidopsis flowering MKYIHILILFKSRSTYKLSTNHI SEQ ID NO: 15 thaliana AtmiPEP161miR161 Arabidopsis  PPR gene family MKIPLFLPKL SEQ ID NO: 16 (10/1199)thaliana DCL1 gene,  AtmiPEP162a1 miR162a Arabidopsis  involvedMVSGQEDSWLKLSSLCFLFLSLLDSLI SEQ ID NO: 17 (4.03/3045) thalianain development AtmiPEP162b1 miR162b Arabidopsis MFLLIFLRLIMICVCSSTDFLRSVNYFC SEQ ID NO: 18 (5.71/4114) thaliana LFIYDLAtmiPEP163-1 miR163 Arabidopsis  SAMT gene family, MSTTQEHRSSEQ ID NO: 19 (6.5/1076) thaliana involved in the AtmiPEP163-2 miR163Arabidopsis  production of MILKCWSSRFLRVSPYQNAHSLSLG SEQ ID NO: 20thaliana secondary metabolites AlmiPEP164a1 miR164a Arabidopsis NAC gene family, MPLAVIRQGIVWP SEQ ID NO: 21 lyrata involved in root, AlmiPEP164a2 miR164a Arabidopsis  foliar and floralMPSWHDMVLLPYVKHTHANTRHIT SEQ ID NO: 22 lyrata devlopment AlmiPEP164a3miR164a Arabidopsis  MTWFFCLT SEQ ID NO: 23 lyrata AtmiPEP164a1 miR164aArabidopsis  MPSWHGMVLLPYVKHTHASTHTHTH SEQ ID NO: 24 (7.05/4256)thaliana NIYGCACELVFH AtmiPEP164a2 miR164a Arabidopsis MAWYGSFALRKTHSRQHTHTHT SEQ ID NO: 25 thaliana AtmiPEP164a3 miR164aArabidopsis  MVWFFCLT SEQ ID NO: 26 thaliana BrmiPEP164a1 miR164aBrassica rapa MMIILWK SEQ ID NO: 27 BrmiPEP164a2 miR164a Brassica rapaMLWAKLVSFSTLHSLVFLLSPSFA SEQ ID NO: 28 BrmiPEP164a3 miR164aBrassica rapa MPSWHGIVILPFVKHTHANIHYSYSC SEQ ID NO: 29 VCI CpmiPEP164a1miR164a Carica papaya MIACHPYLPFPLFLSLTFYSIFFSPSPPS SEQ ID NO: 30 PSLPLCpmiPEP164a2 miR164a Carica papaya MPSLLAFSPFPFSNILLNLLLPLPPFPLSSEQ ID NO: 31 CrmiPEP164a1 miR164a Capsella rubella MELKGLRTWQLLDKVSEQ ID NO: 32 CrmiPEP164a2 miR164a Capsella rubella MPSWHGMACFYCLTSEQ ID NO: 33 CrmiPEP164a3 miR164a Capsella rubellaMAWHGMFLLPYVKHTHANTYSLYM SEQ ID NO: 34 GrmiPEP164a1 miR164a Gossypium MMRSRILKFQYRFGMGIGGRKQLKN SEQ ID NO: 35 raimondii QLCQIQGRISGrmiPEP164a2 miR164a Gossypium  MSNSRSYQLK SEQ ID NO: 36 raimondiiGrmiPEP164a3 miR164a Gossypium  MNEDLEISTRKRTPQLC SEQ ID NO: 37raimondii MtmiPEP164a1 miR164a Medicago  MPKFDIFFYIFV SEQ ID NO: 38truncatula MtmiPEP164a2 miR164a Medicago  MSYISLSPKLLPINTKPFPWLVQFNFYSEQ ID NO: 39 truncatula FSSNTKCNKLHFLGEKLLVGEAGHVQILFLIHSLIMHINIFCTCSPSPTRLPHPSL OsmiPEP164a1 miR164a Oryza sativaMQTHSNTPQSTYSLSLSLSE SEQ ID NO: 40 OsmiPEP164a2 miR164a Oryza sativaMCVCDINMHSMLMLL SEQ ID NO: 41 AlmiPEP165a miR165a Arabidopsis HD-ZIPIII gene MRIKLFQLRGMLSGSRILYIYTCVC SEQ ID NO: 42 lyratafamily, involved AtmiPEP165a miR165a Arabidopsis  in vascular, root,MRVKLFQLRGMLSGSRIL SEQ ID NO: 43 (12.3/2105) thaliana foliar and floralBcmiPEP165a miR165a Brassica  development, and MRMKLFQLRGMLSGSRILYIHKYVYSEQ ID NO: 44 carinata nodulation MLIQVFDHICI BjmiPEP165a miR165aBrassica  MRMKLFQLRGMLSGSRILYIHKYVYI SEQ ID NO: 45 juncea C BnmiPEP165amiR165a Brassica napus MRMKLFQLRGMLSGSRILYIHKYVY SEQ ID NO: 46MIIQVFDHICI BomiPEP165a miR165a Brassica  MRMKLFQLRGMLSGSRILYIHKYVYSEQ ID NO: 47 oleracea MLIQVFDHICI BrmiPEP165a miR165a Brassica rapaMRMKLFQLRGMLSGSRILYIHKYVYIC SEQ ID NO: 48 AtmiPEP166a miR166aArabidopsis  MLDLFRSNNRIEPSDFRFD SEQ ID NO: 49 (4.68/2372) thalianaAtmiPEP166b miR166b Arabidopsis  MRDR SEQ ID NO: 50 (9.35/576) thalianaAtmiPEP167a miR167a Arabidopsis  ARF gene family, MNRKISLSLSSEQ ID NO: 51 (11/1148) thaliana involved in root  AtmiPEP167b1 miR167bArabidopsis  and floral MMGCFVGF SEQ ID NO: 52 (5.27/891) thalianadevelopment AtmiPEP167b2 miR167b Arabidopsis  MQEETYEG SEQ ID NO: 53thaliana AtmiPEP169c1 miR169c Arabidopsis  gene family ofMPHTNLKDLFIFSPNVFFSFAIYLHNS SEQ ID NO: 54 (9.3/7110) thalianaCCAAT-bing factor, WNKNYIHKRENFHNTSFALIFFFSSIM involved in SINYGAtmiPEP169c2 miR169c Arabidopsis  nodulation, MFFFRLLFISTILGTKTTFTNERIFTTPL SEQ ID NO: 55 thaliana drought  LLSFFFFRPLAtmiPEP1691 miR1691 Arabidopsis  resistance,  MRHKES SEQ ID NO: 56(8.52/786) thaliana resistance to  nitrogen  deficiency AtmiPEP171a1miR171a Arabidopsis  GRAS gene family, MNLLKKERQRRRQRSIGSHCIASLVLSEQ ID NO: 57 (11.05/4057) thaliana involved in floral KDGYMKKIAtmiPEP171b miR171b Arabidopsis  foliar, and root MVLSGKLTFSEQ ID NO: 58 (8.5/995) thaliana development MtmiPEP171b1 miR171bMedicago  my corrhization MLLHRLSKFCKIERDIVYIS SEQ ID NO: 59 truncatulanodulation MtmiPEP171b2 miR171b Medicago  development, MKIEESEQ ID NO: 60 truncatula ZmmiPEP171b miR171b Zea maysMHLPSTPSRPPPQHTSLSFLGKEMTKG SEQ ID NO: 61 TTTACFG AtmiPEP171c1 miR171cArabidopsis  MLSLSHFHIC SEQ ID NO: 62 (6.68/1187) thaliana MtmiPEP171emiR171e Medicago  MMVFGKPKKAMLVRFNPKTDLHV SEQ ID NO: 63 truncatulaMtmiPEP171h miR171h Medicago  MASAAKVYMA SEQ ID NO: 64 truncatulaAtmiPEP172a1 miR172a Arabidopsis  AP2 gene family, MASKIW SEQ ID NO: 65(8.5/734) thaliana involved in floral AtmiPEP172a3 miR172a Arabidopsis devolpment MVRFQLSIRD SEQ ID NO: 66 thaliana AtmiPEP172b1 miR172bArabidopsis  MCTYYYLINKYF SEQ ID NO: 67 (7.9/1621) thaliana AtmiPEP172c1miR172c Arabidopsis  MFPAKWCRLES SEQ ID NO: 68 (7.98/1367) thalianaAtmiPEP172e1 miR172e Arabidopsis  MGSLSLFKSQLEILMLLLSLSK SEQ ID NO: 69(8.35/2452) thaliana AtmiPEP172e2 miR172e Arabidopsis MSVYIHVPISLNCFSPKSSC SEQ ID NO: 70 thaliana AtmiPEP172e3 miR172eArabidopsis  MGVPNFRPRNR SEQ ID NO: 71 thaliana AcmiPEP319a1 miR319aArabidopsis  TCP gene family MRSRVSFFFFKIMLFRLLGYRSM SEQ ID NO: 72cebennensis involved in floral AcmiPEP319a2 miR319a Arabidopsis and foliar MHTYIHTISNISSIFFCSKRSFSPFTYIRI SEQ ID NO: 73 cebennensisdevlopment IVVIDPFRIALTFR AhmiPEP319a miR319a Arabidopsis MRSRVSLFLSFSSNFAAYSPRS SEQ ID NO: 74 halleri AlmiPEP319a miR319aArabidopsis  MHTYIPSSSFPISNISSVFFCYKRSFSPY SEQ ID NO: 75 lyrataTYIRIIVVIDPFRIALTFR AtmiPEP319a1 miR319a Arabidopsis MNIHTYHHLLFPSLVFHQSSDVPNALS SEQ ID NO: 76 (6.56/5917)  thalianaLHIHTYEYIIVVIDPFRITLAFR AtmiPEP319a2 miR319a Arabidopsis MFQTLYLFIYIHTNILLLS SEQ ID NO: 77 thaliana BrmiPEP319a miR319aBrassica rapa MFKLYFSAILSTQYMHTYHHRIALIFL SEQ ID NO: 78SILYPSTNYLMSPILNPT CpmiPEP319a miR319a Carica papayaMKIKLGFSLIKIIILLDKNS SEQ ID NO: 79 CrmiPEP319a miR319a Capsella rubellaMHPHTYIHIP SSSFLISSFCL SEQ ID NO: 80 EgmiPEP319a miR319a Eucalyptus MKHIQRWRYGETSGRQGDWKRLEIK SEQ ID NO: 81 grandis VHSNPSLKVKKNTNNFSSSLGrmiPEP319a miR319a Gossypium  MIHFNLSQWRAIMANFHLTYSFLFGC SEQ ID NO: 82raimondii VL MtmiPEP319a miR319a Medicago  MHVYLELFMVIKGLGFLLLVKSEQ ID NO: 83 truncatula OsmiPEP319a miR319a Oryza sativaMEMIQRPCLILKFFFKLSTLYIP SEQ ID NO: 84 PpmiPEP319a miR319a PhyscomitrellaMFHRRRSSVLLPPFGQTQPNPRCLPDL SEQ ID NO: 85 patens RFPSCFTPCTAThmiPEP319a1 miR319a Thellungiella MTICKVSKACFYAGKIENSRLIKKIGIPSEQ ID NO: 86 halophila KREGAPFSPIRENQ ThmiPEP319a2 miR319aThellungiella MEIQIKKKNLYIMNTQKLPNLYIYIYK SEQ ID NO: 87 halophilaYVFIKLMVVE AtmiPEP319b1 miR319b Arabidopsis MVPQINLWSSRVILKIRIDSSTHREEDSEQ ID NO: 88 (8.04/5120)  thaliana HCIQNHKHGLSFIFSF AtmiPEP394a1miR394a Arabidopsis F-box gene family, MSLQFYERVSFKNTVK SEQ ID NO: 89(9.7/1977) thaliana involved in foliar development anddrought resistance AtmiPEP395c1 miR395c Arabidopsis Family of the APS MTEQEEESQMST SEQ ID NO: 90 (3.58/1429) thaliana and AST genes, AtmiPEP395e1 miR395e Arabidopsis involved in  MYLQYIDNVISIYSNNRRVGRMFSRVSEQ ID NO: 91 (9.98/4700) thaliana germination and  PLSTSLEIQFFIKsulphur metabolism AtmiPEP397b1 miR397b Arabidopsis Family of the MSKEIFFSPGFE SEQ ID NO: 92 (4.53/1418) thaliana genes of laccases, involved in copper metabolism, their  overexpression improves growthAtmiPEP398c1 miR398c Arabidopsis CSD gene family,MRTHEQSTAITTLRHCYSSRFMCSQV SEQ ID NO: 93 thaliana involved in copperTPAELFLYRPCFINAVAR metabolism, its overexpression improves growthAtmiPEP399b miR399b Arabidopsis PHO2 gene family, MKRNM SEQ ID NO: 94(11/678) thaliana involved in AtmiPEP399d1 miR399d Arabidopsisphosphorus MQCEI SEQ ID NO: 95 (4/622) thaliana metabolism AtmiPEP403miR403 Arabidopsis AGO gene family MFCA SEQ ID NO: 96 (5.27470) thalianaAtmiPEP447a1 miR447a Arabidopsis Family of the  MVMAHH SEQ ID NO: 97(6.69/724) thaliana genes of  AtmiPEP447a2 miR447a Arabidopsisphosphoglycerate MMKPRWNCSLYGITEWTNNQNQKSK SEQ ID NO: 98 thaliana kinaseRKGRRKTQIWRIGDRLDTVECITLML SAY AtmiPEP447b1 miR447b ArabidopsisMLLIIVELVL SEQ ID NO: 99 (4/1155) thaliana AtmiPEP447b2 miR447bArabidopsis  MLCFNFRCVRRFAE SEQ ID NO: 100  thaliana AtmiPEP447c miR447cArabidopsis MYTYQLDNSFSWFLCTRFCLYRYFLF SEQ ID NO: 101 thalianaNFRCFRRFSE DmmiPEP1a miR1 Drosophila Muscular MWREVCAQKSQTKRRNFITGNQRRNSEQ ID NO: 102 melanogaster differentiation KTKANRKAETKQQKVYEFFVQARERCKTRKKHEKKTLKKTKKIQNRYRAV SENEWGKGFPSHI DmmiPEP1b miR1 DrosophilaMuscular MRTKKSNKKAQFYYGQPTTKQNKSQ SEQ ID NO: 103 melanogasterdifferentiation PKSRNKAAKSL DmmiPEP8 miR8 Drosophila GrowthMEPGFVFVLFPTHLSTQHTQREKSILV SEQ ID NO: 104 melanogasterMGLNLQSAKQSDKQNSKERKKNTQI NSQRIPYRQGGQCSKVLSP HsmiPEP155 miR155Homo sapiens inflammation MEMALMVAQTRKGKSVV SEQ ID NO: 355 AtmiPEP157cmiR157c Arabidopsis SPL gene family, MMLHITHRFESDVGC SEQ ID NO: 375(5.95/1776) thaliana involved in AtmiPEP157d miR157d Arabidopsisdevelopment of the MLYV SEQ ID NO: 376 (5.27/524) thalianastem and flowering AtmiPEP160c miR160c Arabidopsis ARF gene family,MFMRRGLVYNNIYI SEQ ID NO: 377 (9.98/1790) thaliana involved ingermination, development and flowering AtmiPEP164b miR164b ArabidopsisNAC gene family, MMKVCDEQDGEAGHVHY SEQ ID NO: 378 (4.72/1949) thalianainvolved in root,  foliar and floral development AtmiPEP166c miR166cArabidopsis HD-ZIPIII gene MKKRITRINLEEQIKKTLDDSRTRLHS SEQ ID NO: 379(10.42/3407) thaliana family, involved  P in vascular, root,  foliar and floral development and nodulation AtmiPEP166d miR166d ArabidopsisMKKIGSIDSF SEQ ID NO: 380 (8.35/1125) thaliana AtmiPEP169a miR169aArabidopsis  Gene family of MTCRFK SEQ ID NO: 381 (9.5/784) thalianaCCAAT-bing factor, AtmiPEP169h1 miR169h Arabidopsis  MVT (5.28/349)thaliana involved in AtmiPEP169h2 miR169h Arabidopsis  nodulation, MKNENLCGSQG SEQ ID NO: 382 thaliana drought  AtmiPEP169n miR169nArabidopsis  resistance,  MKCMMKKRGLTWRKASCLVAKDDL SEQ ID NO: 383(8.96/5315) thaliana resistance to  PDLFRLHDSISNSCILDYYTF nitrogen deficiency AtmiPEP170 miR170 Arabidopsis  GRAS gene family, MFPRESLSEQ ID NO: 384 (5.75/879) thaliana involved in floral, foliar, and rootdevelopment, mycorrhization, nodulation AtmiPEP396a miR396a Arabidopsis Family of the GRF MTLSVFFHSFLELQNFFRFFFFSFDISY SEQ ID NO: 385 (5.3/3636)thaliana genes involved in  A root development  and cellular proliferation, mycorrhization AtmiPEP399c miR399c Arabidopsis PHO2 gene family, MSLAKGELPCHCFRLNTVYNRFC SEQ ID NO: 386 (8.66/2703)thaliana involved in phosphorus metabolism MtmiPEP169d miR169d Medicago Family NF-YA MVKESFMERLKVR SEQ ID NO: 424 tuncatula genes (or HAP2genes)

TABLE 2 List of the miORFs miPEP Organism Sequence of the miORF SEQ IDAtmiPEP156a1 Arabidopsis thalianaATGTTCTGTTCAATTCAATGCGTCGCCAGACATCTGTTCCCTTTGC SEQ ID NO: 105ATGTAAGAGAGATAAAGAAAGCGACAAGAGCCATAAAGAAAGG TAA AtmiPEP156a2Arabidopsis thaliana ATGCGTCGCCAGACATCTGTTCCCTTTGCATGTAAGAGAGATAAASEQ ID NO: 106 GAAAGCGACAAGAGCCATAAAGAAAGGTAA AtmiPEP156a3Arabidopsis thaliana ATGGTTATGTTTTTTCTCGATTTAGACAAAAACCCTAGATTTGATCSEQ ID NO: 107 TTCTAAAGGGTCTCAAATGGAATCTCTTCTCTTCTCATATCTCTCCCTCTCTCCCTCCCTCTCTTTGA AtmiPEP156c1 Arabidopsis thalianaATGAAGGACAACTTTCCTCTTCTCCTTCGGTTATAA SEQ ID NO: 108 AtmiPEP156c2Arabidopsis thaliana ATGAGTGATGACTGA SEQ ID NO: 109 AtmiPEP156e1Arabidopsis thaliana ATGATATATATAAATAAATATGGGTCGATATCGGCTGTGGAGGACSEQ ID NO: 110 GACTAG AtmiPEP156f1 Arabidopsis thaliana ATGAGCCAAAGATAASEQ ID NO: 111 AlmiPEP159a Arabidopsis lyrataATGACGTGTCCTCTTCTCTCTCTCTCTTTCCTTCTCTCTAAGTATAT SEQ ID NO: 112 TTAGAtmiPEP159a1 Arabidopsis thalianaATGACGTGGCCTCTTCTCTCTCTCTCTTTCCTTCTCTCTAAGTATGT SEQ ID NO: 113 TTAGCrmiPEP159a Capsella rubellaATGACGTGTACTCTCTCTGCTCTATCTCTCTCTCTAAATATGTTTA SEQ ID NO: 114 GGGTTAAAtmiPEP159b1 Arabidopsis thaliana ATGTTTTATCTTTCATAA SEQ ID NO: 115AtmiPEP159b2 Arabidopsis thalianaATGGTTAATACTAGTAGCTTTTTCATTTCAAGTTTTATCCTTCCAT SEQ ID NO: 116TGGTTCTTTCTGAGTCAAATTGTCTCCTGTTTCGAACCATATATAA GTTTTCAATGGTTTTGTATTAAAtmiPEP160a1 Arabidopsis thalianaATGTTTTGTTTGTTGATTCCCATCTTCTCTTTTGTCTTTTCACCAAA SEQ ID NO: 117TCGTCATTTAAGGCTTCAAGAACAGTAA AtmiPEP160b1 Arabidopsis thalianaATGTTTTCCCCTCAATGA SEQ ID NO: 118 AtmiPEP160b2 Arabidopsis thalianaATGAAATACATACACATTTTGATTTTATTTAAATCAAGATCGACG SEQ ID NO: 119TATAAGCTATCCACCAATCATATTTAA AtmiPEP161 Arabidopsis thalianaATGAAAATTCCATTGTTTCTGCCGAAGCTTTGA SEQ ID NO: 120 AtmiPEP162a1Arabidopsis thaliana ATGGTATCTGGTCAAGAAGATTCCTGGTTAAAACTTTCATCTCTCTSEQ ID NO: 121 GTTTCCTTTTTCTTTCTTTGTTGGATTCATTAATTTGA AtmiPEP162b1Arabidopsis thaliana ATGTTTCTTTTAATCTTTTTGAGATTAATAATGATTTGTGTTTGTTCSEQ ID NO: 122 ATCAACCGATTTTCTCAGATCTGTCAATTATTTTTGTTTATTTATTTATGATTTATGA AtmiPEP163-1 Arabidopsis thalianaATGTCCACTACTCAAGAGCATAGGTCTTGA SEQ ID NO: 123 AtmiPEP163-2Arabidopsis thaliana ATGATACTAAAGTGCTGGAGTTCCCGGTTCCTGAGAGTGAGTCCASEQ ID NO: 124 TATCAAAATGCGCATTCGTTATCACTTGGTTGA AlmiPEP164a1Arabidopsis lyrata ATGCCCTTAGCAGTTATTAGACAAGGGATTGTTTGGCCCTAGSEQ ID NO: 125 AlmiPEP164a2 Arabidopsis lyrataATGCCATCATGGCATGACATGGTTCTTTTGCCTTACGTAAAACAC SEQ ID NO: 126ACTCACGCCAACACACGCCACATAACATAA AlmiPEP164a3 Arabidopsis lyrataATGACATGGTTCTTTTGCCTTACGTAA SEQ ID NO: 127 AtmiPEP164a1Arabidopsis thaliana ATGCCATCATGGCATGGTATGGTTCTTTTGCCTTACGTAAAACACSEQ ID NO: 128 ACTCACGCCAGCACACACACACACACACATAACATATACGGATGTGCGTGTGAGCTAGTCTTCCATTAA AtmiPEP164a2 Arabidopsis thalianaATGGCATGGTATGGTTCTTTTGCCTTACGTAAAACACACTCACGC SEQ ID NO: 129CAGCACACACACACACACACATAA AtmiPEP164a3 Arabidopsis thalianaATGGTATGGTTCTTTTGCCTTACGTAA SEQ ID NO: 130 BrmiPEP164a1 Brassica rapaATGATGATAATTTTGTGGAAATAA SEQ ID NO: 131 BrmiPEP164a2 Brassica rapaATGCTTTGGGCCAAGCTAGTTTCTTTTAGCACTCTTCACTCACTAG SEQ ID NO: 132TTTTTCTTCTCAGCCCTTCTTTTGCGTGA BrmiPEP164a3 Brassica rapaATGCCATCATGGCATGGCATTGTCATTTTGCCTTTCGTAAAACAC SEQ ID NO: 133ACTCACGCCAACATACATTATTCATATTCATGTGTATGTATATGAATGCCATCATGGCATATGCCATCATGGCAT CpmiPEP164a1 Carica papayaATGATTGCATGCCATCCCTACTTGCCTTTTCCCCTTTTCCTTTCTCT SEQ ID NO: 134AACATTTTACTCAATCTTCTTCTCCCCCTCCCCCCCTTCCCCCTCTC TGCCATTATAA CpmiPEP164a2Carica papaya ATGCCATCCCTACTTGCCTTTTCCCCTTTTCCTTTCTCTAACATTTTSEQ ID NO: 135 ACTCAATCTTCTTCTCCCCCTCCCCCCCTTCCCCCTCTCTGCCATTATAACCATAATTAAACCTCTCTCCCTCTCTCTCCCTCTCTCTCTCTCT CTCTCTGGGTTCTCAGTATAACrmiPEP164a1 Capsella rubellaATGGAATTAAAAGGTTTGAGAACTTGGCAGTTATTATTAGACAAGGT SEQ ID NO: 136 ATAGCrmiPEP164a2 Capsella rubellaATGCCATCATGGCATGGCATGGCATGTTTCTATTGCCTTACGTAA SEQ ID NO: 137CrmiPEP164a3 Capsella rubellaATGGCATGGCATGTTTCTATTGCCTTACGTAAAACACACTCACGC SEQ ID NO: 138CAACACATACTCACTATACATGTAAATAAGTATGTGCGCGTGTGA GrmiPEP164a1Gossypium raimondii ATGATGAGATCAAGAATTTTAAAGTTTCAATATAGATTTGGCATGSEQ ID NO: 139 GGTATTGGCGGCAGAAAGCAATTAAAAAACCAGTTATGTCAAATTCAAGGTCGTATCAGTTAA GrmiPEP164a2 Gossypium raimondiiATGTCAAATTCAAGGTCGTATCAGTTAAAATGA SEQ ID NO: 140 GrmiPEP164a3Gossypium raimondii ATGAATGAAGATTTAGAAATTTCAACAAGGAAGAGGACCCCACASEQ ID NO: 141 GCTTTGTTAA MtmiPEP164a1 Medicago truncatulaATGCCCAAATTTGATATTTTTTTTTATATATTTGTATAG SEQ ID NO: 142 MtmiPEP164a2Medicago truncatula ATGTCATATATCTCTCTCTCTCCTAAGTTGCTACCTATAAATACTASEQ ID NO: 143 AGCCTTTCCCTTGGTTGGTTCAATTCAACTTCTACTTCTCATCAAACACAAAGTGCAATAAGCTTCATTTCCTGGGTGAGAAGCTCCTTGTTGGAGAAGCAGGGCACGTGCAAATCCTCTTTCTGATTCATTCTCTCATAATGCATATCAATATCTTTTGCACGTGCTCCCCTTCTCCAACT AGG OsmiPEP164a1Oryza sativa ATGCAAACCCACTCCAACACTCCACAATCCACATACTCTCTCTCTSEQ ID NO: 144 CTCTCTCTCTCTGAGTAG OsmiPEP164a2 Oryza sativaATGTGTGTGTGTGATATCAATATGCATTCGATGTTGATGCTACTGT SEQ ID NO: 145 AGAlmiPEP165a Arabidopsis lyrataATGAGAATTAAGCTATTTCAGTTGAGGGGAATGTTGTCTGGATCG SEQ ID NO: 146AGGATATTATACATATATACATGTGTATGTTGA AtmiPEP165a Arabidopsis thalianaATGAGGGTTAAGCTATTTCAGTTGAGGGGAATGTTGTCTGGATCG SEQ ID NO: 147AGGATATTATAG BcmiPEP165a Brassica carinataATGAGAATGAAGCTATTTCAGTTGAGGGGAATGTTGTCTGGATCG SEQ ID NO: 148AGGATATTATATATACACAAATACGTATATATGTTAATACAAGTG TTTGATCATATATGTATATAGBjmiPEP165a Brassica junceaATGAGAATGAAGCTATTTCAGTTGAGGGGAATGTTGTCTGGATCG SEQ ID NO: 149AGGATATTATATATACACAAATATGTATATATATGTTAA BnmiPEP165a Brassica napusATGAGAATGAAGCTATTTCAGTTGAGGGGAATGTTGTCTGGATCG SEQ ID NO: 150AGGATATTATATATACACAAATACGTATATATGATAATACAAGTG TTTGATCATATATGTATATAGBomiPEP165a Brassica oleraceaATGAGAATGAAGCTATTTCAGTTGAGGGGAATGTTGTCTGGATCG SEQ ID NO: 151AGGATATTATATATACACAAGTACGTATATATGTTAATACAAGTG TTTGATCATATATGTATATAGBrmiPEP165a Brassica rapa ATGAGAATGAAGCTATTTCAGTTGAGGGGAATGTTGTCTGGATCGSEQ ID NO: 152 AGGATATTATATATACACAAATATGTATATATATGTTAA AtmiPEP166aArabidopsis thaliana ATGTTGGATCTCTTTCGATCTAACAATCGAATTGAACCTTCAGATTSEQ ID NO: 153 TCAGATTTGATTAG AtmiPEP166b Arabidopsis thalianaATGAGAGATAGATAA SEQ ID NO: 154 AtmiPEP167a Arabidopsis thalianaATGAACAGAAAAATCTCTCTTTCTCTTTCTTGA SEQ ID NO: 155 AtmiPEP167b1Arabidopsis thaliana ATGATGGGTTGTTTTGTGGGATTTTAA SEQ ID NO: 156AtmiPEP167b2 Arabidopsis thaliana ATGCAGGAGGAAACATATGAGGGGTGASEQ ID NO: 157 AtmiPEP169c1 Arabidopsis thalianaATGCCACATACAAACTTGAAAGATCTCTTCATCTTTTCTCCAAATG SEQ ID NO: 158TTTTTTTTTCGTTTGCTATTTATCTCCACAATTCTTGGAACAAAAACTACATTCACAAACGAGAGAATTTTCACAACACCTCTTTTGCTCTCATTTTTTTTTTTTCGTCCATTATGAGTATTAATTATGGTTAG AtmiPEP169c2Arabidopsis thaliana ATGTTTTTTTTTCGTTTGCTATTTATCTCCACAATTCTTGGAACAASEQ ID NO: 159 AAACTACATTCACAAACGAGAGAATTTTCACAACACCTCTTTTGCTCTCATTTTTTTTTTTTCGTCCATTATGA AtmiPEP16911 Arabidopsis thalianaATGAGACATAAAGAGAGTTAA SEQ ID NO: 160 AtmiPEP171a1 Arabidopsis thalianaATGAACCTCCTCAAGAAGGAAAGACAGAGGAGGAGACAAAGAA SEQ ID NO: 161GTATAGGTTCACATTGCATAGCCAGTTTAGTTTTGAAGGATGGAT ATATGAAAAAAATATGAAtmiPEP171b Arabidopsis thaliana ATGGTTCTCTCCGGTAAATTAACATTTTAGSEQ ID NO: 162 MtmiPEP171b1 Medicago truncatulaATGCTTCTTCATAGGCTCTCCAAATTTTGCAAAATTGAAAGAGAC SEQ ID NO: 163ATAGTATATATATCTTAG MtmiPEP171b2 Medicago truncatula ATGAAGATTGAAGAGTAASEQ ID NO: 164 ZmmiPEP171b Zea maysATGCATCTGCCTTCAACTCCCTCTCGCCCCCCACCCCAACACACAT SEQ ID NO: 165CTCTCTCTTTTCTAGGGAAGGAAATGACGAAGGGGACGACGACG GCATGCTTCGGCTAGAtmiPEP171c1 Arabidopsis thaliana ATGTTGTCTCTTTCTCATTTTCATATCTGCTAASEQ ID NO: 166 MtmiPEP171e Medicago truncatulaATGATGGTGTTTGGGAAGCCGAAAAAAGCGATGTTGGTGAGGTT SEQ ID NO: 167CAATCCGAAGACGGATTTACATGTATAG MtmiPEP171h Medicago truncatulaATGGCTTCAGCTGCAAAAGTATACATGGCGTGA SEQ ID NO: 168 AtmiPEP172a1Arabidopsis thaliana ATGGCTTCCAAGATCTGGTAA SEQ ID NO: 169 AtmiPEP172a3Arabidopsis thaliana ATGGTTAGGTTCCAACTAAGTATACGAGATTAA SEQ ID NO: 170AtmiPEP172b1 Arabidopsis thalianaATGTGTACGTACTATTATCTCATAAATAAATATTTTTAA SEQ ID NO: 171 AtmiPEP172c1Arabidopsis thaliana ATGTTTCCAGCAAAATGGTGCCGTCTTGAGTCTTGA SEQ ID NO: 172AtmiPEP172e1 Arabidopsis thalianaATGGGATCTCTCTCTTTATTTAAAAGTCAATTAGAGATCTTGATGC SEQ ID NO: 173TACTTCTGTCCCTTTCCAAGTGA AtmiPEP172e2 Arabidopsis thalianaATGAGTGTATATATTCATGTACCTATCTCTCTCAATTGCTTCTCAC SEQ ID NO: 174CAAAATCATCTTGCTGA AtmiPEP172e3 Arabidopsis thalianaATGGGAGTTCCCAACTTTAGACCTCGAAACCGATAA SEQ ID NO: 175 AcmiPEP319a1Arabidopsis cebennensis TAGAGATCTAGGGTTTCTTTCTTTTTCTTCAAAATCATGCTTTTTCSEQ ID NO: 176 GCTTGCTAGGTTATAGATCCATGTAA AcmiPEP319a2Arabidopsis cebennensis ATGCATACATACATACATACCATCTCTAATATTTCATCAATCTTCTSEQ ID NO: 177 TTTGTTCCAAACGCCTTTCTCTCCATTTACATACATACGAATCATTGTTGTCATAGATCCGTTTAGAATTGCTTTAACTTTTAGATGA AhmiPEP319aArabidopsis halleri ATGAGATCTAGGGTTTCTTTGTTTCTTTCGTTTTCTTCAAATTTTGCSEQ ID NO: 178 TGCATATTCTCCAAGATCATGA AlmiPEP319a Arabidopsis lyrataATGCATACATACATACCATCATCATCTTTTCCCATCTCTAATATTT SEQ ID NO: 179CATCAGTCTTCTTTTGTTACAAACGCTCTTTCTCGCCATATACATACATAAGAATCATTGTTGTCATAGATCCGTTTAGAATTGCTTTAACT TTTAGATGA AtmiPEP319a1Arabidopsis thaliana ATGAATATACATACATACCATCATCTTCTTTTCCCATCTCTAGTTTSEQ ID NO: 180 TTCATCAATCTTCTGATGTTCCAAACGCTCTATCTCTTCATATACATACATACGAATATATTATTGTTGTCATAGATCCATTTAGAATCACT TTAGCTTTTAGATGAAtmiPEP319a2 Arabidopsis thalianaATGTTCCAAACGCTCTATCTCTTCATATACATACATACGAATATAT SEQ ID NO: 181TATTGTTGTCATAG BrmiPEP319a Brassica rapaATGTTTAAGCTCTACTTCTCAGCAATTCTCTCCACCCAATACATGC SEQ ID NO: 182ATACATACCATCATCGTATCGCTCTAATTTTTCTATCAATCTTGTATCCTTCCACAAATTATCTTATGTCTCCCATTTTAAATCCTACATAG CpmiPEP319a Carica papayaATGAAGATTAAATTAGGTTTTAGTCTTATTAAGATTATTATATTAC SEQ ID NO: 183TAGACAAAAACAGTTAA CrmiPEP319a Capsella rubellaATGCATCCACATACATACATACATATACCATCATCTTCTTTTCTCA SEQ ID NO: 184TCTCTAGTTTTTGTTTATAA EgmiPEP319a Eucalyptus grandisATGAAGCATATTCAAAGGTGGAGATATGGGGAGACTTCCGGAAG SEQ ID NO: 185GCAAGGGGATTGGAAAAGGCTCGAGATCAAAGTGCATAGCAACCCTTCGCTAAAGGTGAAAAAGAATACGAATAACTTCAGTAGCTCAC TTTAA GrmiPEP319aGossypium raimondii ATGATCCATTTCAACCTGTCACAGTGGAGAGCAATTTGTATGGCTSEQ ID NO: 186 AATTTCCATCTCACCTATTCTTTTCTGTTTGGGGTTCTCTAG MtmiPEP319aMedicago truncatula ATGCATGTATATCTTGAATTGTTTATGGTAATAAAGGGGTTAGGASEQ ID NO: 187 TTTCTCCTTTTGGTGAAGTGA OsmiPEP319a Oryza sativaATGGAAATGATACAAAGGCCGTGTTTAATTTTAAAATTTTTTTTCA SEQ ID NO: 188AACTTTCAACACTTTACATCCCATAA PpmiPEP319a Physcomitrella patensATGTTCCACCGTCGGAGATCCTCGGTGCTGCTACCCCCGTTCGGC SEQ ID NO: 189CAAACCCAACCCAACCCTAGGTGTCTGCCGGACCTCCGCTTCCCCTCCTGCTTCACCCCCTGCACCGCTTAA ThmiPEP319a1 Thellungiella halophilaATGACGATATGTAAAGTAAGCAAGGCATGTTTTTATGCAGGGAA SEQ ID NO: 190GATTGAAAATTCAAGATTAATCAAGAAAATTGGAATACCAAAAAGAGAGGGAGCTCCCTTCAGTCCAATCAGAGAGAATCAATGA ThmiPEP319a2Thellungiella halophila ATGGAGATTCAAATTAAAAAGAAAAACTTATATATAATGAATACSEQ ID NO: 191 ACAAAAGCTACCTAATCTGTATATATATATATATAAATATGTCTTCATTAAATTAATGGTCGTGGAATAG AtmiPEP319b1 Arabidopsis thalianaATGGTACCTCAAATTAATCTATGGTCATCTAGGGTTATCTTGAAG SEQ ID NO: 192ATTAGAATTGATTCTAGCACGCACAGAGAGGAAGATCATTGCATCCAGAATCACAAACATG3GCCTATCTTTTATCTTTTCTTTTTGA AtmiPEP394a1Arabidopsis thaliana ATGTCTCTCCAATTTTATGAGAGGGTTTCCTTCAAGAACACAGTASEQ ID NO: 193 AAATAG AtmiPEP395c1 Arabidopsis thalianaATGACAGAGCAAGAAGAAGAAAGTCAAATGTCCACATGA SEQ ID NO: 194 AtmiPEP395e1Arabidopsis thaliana ATGTATCTACAATATATTGATAATGTAATATCTATATATTCAAACSEQ ID NO: 195 AATCGTCGTGTTGGTCGGATGTTTTCTAGAGTTCCTCTGAGCACTTCATTGGAGATACAATTTTTTATAAAATAG AtmiPEP397b1 Arabidopsis thalianaATGAGCAAGGAGATATTTTTTTCCCCTGGGTTTGAATGA SEQ ID NO: 196 AtmiPEP398c1Arabidopsis thaliana ATGAGAACACACGAGCAATCAACGGCTATAACGACGCTACGTCASEQ ID NO: 197 TTGTTACAGCTCTCGTTTCATGTGTTCTCAGGTCACCCCTGCTGAGCTCTTTCTCTACCGTCCATGTTTTATCAACGCCGTGGCCCGTG AtmiPEP399bArabidopsis thaliana ATGAAGAGAAACATGTAA SEQ ID NO: 198 AtmiPEP399d1Arabidopsis thaliana ATGCAATGTGAAATATGA SEQ ID NO: 199 AtmiPEP403Arabidopsis thaliana ATGTTTTGTGCTTGA SEQ ID NO: 200 AtmiPEP447a1Arabidopsis thaliana ATGGTCATGGCTCATCATTAG SEQ ID NO: 201 AtmiPEP447a2Arabidopsis thaliana ATGATGAAACCTCGATGGAACTGCTCTCTTTATGGAATCACGGAASEQ ID NO: 202 TGGACAAATAATCAAAATCAGAAATCGAAGCGAAAAGGGAGGAGAAAAACGCAGATTTGGAGGATTGGGGACAGATTAGATACTGTTGAATGCATCACTCTAATGCTATCAGCCTATTAA AtmiPEP447b1 Arabidopsis thalianaATGCTGCTTATCATCGTGGAGTTGGTTCTGTAA SEQ ID NO: 203 AtmiPEP447b2Arabidopsis thaliana ATGCTTTGTTTCAATTTCAGGTGCGTTAGAAGGTTTGCAGAGTAGSEQ ID NO: 204 AtmiPEP447c Arabidopsis thalianaATGTACACCTACCAGCTTGATAACTCTTTTTCGTGGTTTCTGTGTA SEQ ID NO: 205CTCGTTTCTGTTTGTACAGATACTTCTTGTTCAATTTCAGATGCTTT AGAAGGTTTTCGGAGdmmiPEP1a Drosophila melanogasterATGTGGCGCGAAGTATGCGCACAAAAAAGTCAAACAAAAAGGCG SEQ ID NO: 206CAATTTTATTACGGGCAACCAACGACGAAACAAAACAAAAGCCAACCGAAAAGCAGAAACAAAGCAGCAAAAAGTTTATGAATTTTTTGTGCAGGCGCGTGAAAGATGCAAAACGAGAAAAAAACATGAAAAAAAAACATTAAAAAAAACAAAAAAAATCCAAAACAGATACCGAGCTGTATCCGAAAACGAGTGGGGAAAGGGGTTTCCCAGTCACA TATAA DmmiPEP1bDrosophila melanogaster ATGCGCACAAAAAAGTCAAACAAAAAGGCGCAATTTTATTACGGSEQ ID NO: 207 GCAACCAACGACGAAACAAAACAAAAGCCAACCGAAAAGCAGAAACAAAGCAGCAAAAAGTTTATGA DmmiPEP8 Drosophila melanogasterATGGAGCCTGGCTTTGTTTTTGTTTTATTTCCAACCCACTTGAGCA SEQ ID NO: 208CACAGCACACACAGAGAGAAAAATCAATACTCGTTATGGGATTAAATTTACAAAGCGCAAAGCAAAGCGACAAACAAAATTCAAAAGAAAGAAAAAAAAACACTCAAATAAACTCACAAAGAATTCCTTATCGCCAAGGGGGCCAATGTTCTAAGGTTCTTTCGCCTTGA HsmiPEP155 Homo sapiensTGGAGATGGCTCTAATGGTGGCACAAACCAGGAAGGGGAAATCT SEQ ID NO: 356 GTGGTTTAAAtmiPEP157c Arabidopsis thalianaATGATGTTGCATATCACACATAGGTTTGAGAGTGATGTTGGTTGT SEQ ID NO: 387 TGAAtmiPEP157d Arabidopsis thaliana ATGCTGTATGTATAG SEQ ID NO: 388AtmiPEP160c Arabidopsis thalianaATGTTCATGCGTAGAGGTTTGGTATACAACAATATATACATATAA SEQ ID NO: 389 AtmiPEP164bArabidopsis thaliana ATGATGAAGGTGTGTGATGAGCAAGATGGAGAAGCAGGGCACGTSEQ ID NO: 390 GCATTACTAG AtmiPEP166c Arabidopsis thalianaATGAAGAAGAGAATCACTCGAATTAATTTGGAAGAACAAATTAA SEQ ID NO: 391GAAAACCCTAGATGATTCTCGGACCAGGCTTCATTCCCCCTAA AtmiPEP166dArabidopsis thaliana ATGAAGAAGATCGGTAGtATTGATTCATTTTAA SEQ ID NO: 392AtmiPEP169a Arabidopsis thaliana ATGACTTGCCGATTTAAATGA SEQ ID NO: 393AtmiPEP169h1 Arabidopsis thaliana ATGGTGACATGA SEQ ID NO: 394AtmiPEP169h2 Arabidopsis thaliana ATGAAGAATGAGAACTTGTGTGGTAGCCAAGGATGASEQ ID NO: 395 AtmiPEP169n Arabidopsis thalianaATGAAGTGTATGATGAAGAAGAGAGGTCTAACATGGCGGAAAGC SEQ ID NO: 396GTCATGTTTAGTAGCCAAGGATGACTTGCCTGATCTTTTTCGCCTCCACGATTCAATTTCAAATTCATGCATTTTGGATTATTATACCTTTT AA AtmiPEP170Arabidopsis thaliana ATGTTTCCGAGAGAGTCCCTCTGA SEQ ID NO: 397 AtmiPEP396aArabidopsis thaliana ATGACCCTCTCTGTATTCTTCCACAGCTTTCTTGAACTGCAAAACTSEQ ID NO: 398 TCTTCAGATTTTTTTTTTTTTCTTTTGATATCTCTTACGCATAA AtmiPEP399cArabidopsis thaliana ATGTCACTTGCCAAAGGAGAGTTGCCCTGTCACTGCTTCCGCTTASEQ ID NO: 399 AACACAGTCTATAACCGGTTCTGCTAA MtmiPEP169dMedicago truncatula ATGGTCAAAGAGTCTTTCATGGAGAGGTTGAAAGTGAGATGASEQ ID NO: 425

TABLE 3 List of the primary transcripts (pri-miRNAs) miPEP OrganismSequence of the Pri-miRNA SEQ ID AtmiPEP156a1 Arabidopsis ATTCATTGTTCACTCTCAAATCTCAAGTTCATTGCCATTTTTAGGTCTCTC SEQ ID NO: 209AtmiPEP156a2 thaliana TATAAATTCAAATGTTCTGTTCAATTCAATGCGTCGCCAGACATCTGTTCAtmiPEP156a3 CCTTTGCATGTAAGAGAGATAAAGAAAGCGACAAGAGCCATAAAGAAAGGTAAGACTCTTTGAAATAGAGAGAGATAAGGTTTTCTCTTATCTTCTTCTCATCAGATCTTTGTTTCTTTACCCTCTTTCTTTCTTTTTTTTGCTTTTTATGGTTATGTTTTTTCTCGATTTAGACAAAAACCCTAGATTTGATCTTCTAAAGGGTCTCAAATGGAATCTCTTCTCTTCTCATATCTCTCCCTCTCTCCCTCCCTCTCTTTGATTCTTTGTCTTCTCCAGTTAAAACTCAGATCTAACACAAAGCTTAAAAGATTCTCATCGTTTCTTGTTTTCTTTGTTTCATCTTGTAGATCTCTGAAGTTGGACTAATTGTGAATGAAAGAGTTGGGACAAGAGAAACGCAAAGAAACTGACAGAAGAGAGTGAGCACACAAAGGCAATTTGCATATCATTGCACTTGCTTCTCTTGCGTGCTCACTGCTCTTTCTGTCAGATTCCGGTGCT GATCTCTTTAtmiPEP156c1 Arabidopsis CTCTGCCTTTAGTTCTTTCTTTTTTGGTAATATATTTATTTTTCGTTACGAT SEQ ID NO: 210AtmiPEP156c2 thaliana TTGGTCAAAACCCTAGATTTGTTTTCCAAAAGCATATCTGAAAATGAAGGACAACTTTCCTCTTCTCCTTCGGTTATAAATATTCTCTCCGGTTTTGCTTGTTTAACCTAAAAGCCTCAGATCTAACTCCAACACCTTCAAAGTCTGCCTCCTTTCCAATCTTCTTTCTTCTGTTCGATCTCTAATCTCAGAATTTGTGTCGGTAAGGTAAAGGTGATAATGAGTGATGACTGATGAGGGAGTTTTGGGACAAATTTTAAGAGAAACGCATAGAAACTGACAGAAGAGAGTGAGCACACAAAGGCACTTTGCATGTTCGATGCATTTGCTTCTCTTGCGTGCTCACTGCTCT ATCTGTCAGATTCCGGCTAtmiPEP156e1 Arabidopsis TCCCACATCCAAAGATAGAAAGATGTAAGGTCTAGAGTCTTGTTCTTAAT SEQ ID NO: 211thaliana CCCCTAACAGAACAATGATATATATAAATAAATATGGGTCGATATCGGCTGTGGAGGACGACTAGCTACGGTTTCGAGCCTGGTCACATGCGTAGAGTGTGAAAGGTAATTAGGAGGTGACAGAAGAGAGTGAGCACACATGGTGGTTTCTTGCATGCTTTTTTGATTAGGGTTTCATGCTTGAAGCTATGTGTGCTTACTCTCTCTCTGTCACCCCT AtmiPEP156f1 Arabidopsis TCCCACAGCCAATGAGCCAAAGATAAAGAAACACCTATCCTATAATAAT SEQ ID NO: 212thaliana TTAGAGCAATATACCTCCATAATGGAACATCTATATATATAAAGGTATCCGTATATCTCTATATATTATATTCATTGAGTTTAAAGTGGCTAGGGTTTATAGATGTATGTGATATTAAGAGATATGAAACATATTTGTCGACGGTTTGAGTGGTGAGGAATTGATGGTGACAGAAGAGAGTGAGCACACATGGTGGCTTTCTTGCATATTTGAAGGTTCCATGCTTGAAGCTATGTGTGCTCACTCTCTATCCGTCACCCCCTTCTCTCCCTCTCCCTC AlmiPEP159a Arabidopsis AAAAAATGACGTGTCCTCTTCTCTCTCTCTCTTTCCTTCTCTCTAAGTATA SEQ ID NO: 213lyrata TTTAGGGTTAATTATTAGGGTTCTTTATCTCTTTCTTCAGTCTTTGAAGTTTCTTCAATAGCTTTAATTGAAGTGATTTACCTCTCTGGGTGTTTTTAGTATATATATCATGTACATGATCGAATTTCTTTCTATCCAAGTTCTCATCAAACCTTCTCATGTTTTGAAGAGTTAAAGGCTTTATAGTTTGCTTAGGTCAGATCCATAACATACTGTATTTGACAAGTTTCTTTGTCTCACGATAGATCTTGGTCTGACCAAAATGATTTTCTCGAGAAAAAAAAAGATGGAAGTAGAGCTCCTTGAAGTTCAAACGAGAGTTGAGCAGGGTAAAGAAAAGCTGCTAAGCTATGGATCCCATAAGCCCTAATCCTTATAAAGAAAAAAAAGGATTTGGTTATATGGCTTGCATATCTCAGGAGCTTTAACTTGCCCTTTAATGGCTTTTACTCTTCTTTGGATTGAAGGGAGCTCTACATCTTCTTTCACCTTCTCTATTTTTCTTTCTTTATTTTCTCCTCTACAGTAATTTATTTGGATT AtmiPEP159a1 Arabidopsis TTCCAAAACATGACGTGGCCTCTTCTCTCTCTCTCTTTCCTTCTCTCTAAG SEQ ID NO: 214thaliana TATGTTTAGGGTTAATAATTAGGGTTCCTCCTCTCTTTTGTTCTGTCTTTATATCTCCTTCATAGCTCTAATGTAAGAGATTTACCTCTTTTGGTGTTTTTGTTAATCCACGTTCTCATCAAAACTTTCTCATTGTTTTATGAAGAGTTAAAGGTCTTTACAGTTTGCTTATGTCAGATCCATAATATATTTGACAAGATACTTTGTTTTTCGATAGATCTTGATCTGACGATGGAAGTAGAGCTCCTTAAAGTTCAAACATGAGTTGAGCAGGGTAAAGAAAAGCTGCTAAGCTATGGATCCCATAAGCCCTAATCCTTGTAAAGTAAAAAAGGATTTGGTTATATGGATTGCATATCTCAGGAGCTTTAACTTGCCCTTTAATGGCTTTTACTCTTCTTTGGATTGAAGGGAGCTCTACATCTTCTTTCACCTTCTCTATTTTTTATTTTTCTTTATTTCTACTCAACAATTATTTATTCGGATTCATCTTTAATTTTCCGTTATAATTTCTTTTTGGTAAGGATTATTCGCTATAATTTGAGAAT CrmiPEP159a Capsella TTCCAACGAATGACGTGTACTCTCTCTGCTCTATCTCTCTCTCTAAATATG SEQ ID NO: 215rubella TTTAGGGTTAATTAGGGTTCTTCATCTGTCTCTCTCTCTCTCTCTCTTCAGAGTCTTTATAGCTTCTTCCAAGATTTTTAATTGAAAGTAATTTACCTCTTTTGGAGTTCTGTACATATAGAATATCAGGAGTCGTGTTTCTTTTTTATCAAGGTTCTCATCTAACCTTTATAGTATTTTCATTAGTTGATAAAGGTCTTCATAGTTTGCTTAGATCAGATCTTGTCTTCGTCTTTTCGATAGATCTTGTTCTGTCCAATATACAGTGATTTTATTTCGAGAGCAAAAAAGATGAGAGGTAGAGCTCCTTGAAGTTCAAACGAGAGTTTAGCAGGGTAGAGAAAAGCTGCTAAGCTATGGATCCCATAAGCCCTAATCCTTGTTAATGATAAAGGATTTGGTTATATGGCTTGCATATCTCAGGAGCTTTAACTTGCCCTTTAATTGCTTTTACTCTTCTTTGGATTGAAGGGAGCTCTACATCTTCTTTGACTTCTCTCTCTATTAAGTCTTTCTTTATTTTCTTCTCTACAATAGTTGTTTTGGATCGGAAGA TCTTTAAGTTTCCCTTAAtmiPEP159b1 Arabidopsis TTTCACTTTTGTTCTCCTCCTCCCTTTTTTTCTTTTCAGGATTCTTCTTTTCT SEQ ID NO: 216AtmiPEP159b2 thalianaATGTTTTATCTTTCATAATAGATCTGATAATTTTGATTTTTCACTATATATATTATGGTTAATACTAGTAGCTTTTTCATTTCAAGTTTTATCCTTCCATTGGTTCTTTCTGAGTCAAATTGTCTCCTGTTTCGAACCATATATAAGTTTTCAATGGTTTTGTATTAACTCAAGTATTCAACATTATGTCTCTCTTTTTCTTGCTTGGATCTCTAATGCTGTTCATATTTTAAAGCATAGGTTTAGGTTAGATGCATGTAACTGCCAATTAAAAGAAGGTCAAGAGTTTTTTGATTGTATGAATATATGAGTTAGTCAAAGCAGATCCACACGATTATATAGAAAAACAAAGGAAGAAGAAGAGGAAGAGCTCCTTGAAGTTCAATGGAGGGTTTAGCAGGGTGAAGTAAAGCTGCTAAGCTATGGATCCCATAAGCCTTATCAAATTCAATATAATTGATGATAAGGTTTTTTTTATGGATGCCATATCTCAGGAGCTTTCACTTACCCCTTTAATGGCTTCACTCTTCTTTGGATTGAAGGGAGCTCTTCA TCTCTC AtmiPEP160a1Arabidopsis  CATCCCACCCTTAATTGTTTTATATAAACCATTTCTCCTCCTCTCTCCATCSEQ ID NO: 217 thalianaACCTTCAATCTCTCTCGATCTCTCTCTGGATCCCCAATCTCACCTCCATGTTTTGTTTGTTGATTCCCATCTTCTCTTTTGTCTTTTCACCAAATCGTCATTTAAGGCTTCAAGAACAGTAACCCCAATTCCTCCACAAGAGGGAGAGAAAACAAAAGATCTTCCAATTCCATTCTCGTACATGCAAATCACAATCCATGCCATAGATTGTTTCTATTCCTCCTTATTTATTGCTTGTATCTGTTCATGCATGGACCAGGTGGAGAGAGCATTACTTAAAAATAGAATTAGCTATCTGTTTTAGGCGAATTAGTTTCCTTACATAACCATGTATATGTCATGACGCATATACATATGTAGATGTATATGTATTATATATGTATGCCTGGCTCCCTGTATGCCATATGCTGAGCCCATCGAGTATCGATGACCTCCGTGGATGGCGTATGAGGA GCCATGCATATAtmiPEP160b1 Arabidopsis ACTCATAACTCTCCCCAAATTCTTGACCAAAAATATCCGCCACTTTCTCTC SEQ ID NO: 218AtmiPEP160b2 thalianaTGGTTCATGTTTTCCCCTCAATGAAATACATACACATTTTGATTTTATTTAAATCAAGATCGACGTATAAGCTATCCACCAATCATATTTAAGGGTTCCCGTATACATATATACTATATATATATATGGAATAATAGTCGTGCCTGGCTCCCTGTATGCCACAAGAAAACATCGATTTAGTTTCAAAATCGATCACTAGTGGCGTACAGAGTAGTCAAGCATGAC AtmiPEP 161 Arabidopsis CTCTAACTCATCCTTCTCTTCTATGAAAATTCCATTGTTTCTGCCGAAGCT SEQ ID NO: 219thaliana TTGATCAGTACTTCTCTTTTGCTTGATCTCGGTTTTTGACCAGTTTATTGCGTCGATCAATGCATTGAAAGTGACTACATCGGGGTTCCGATTTTTTTTGTTCTTCATATGATGAAGCGGAAACAGTAATCAACCCTGGTTTAGTCACTTTCACTGCATTAATCAATGCATTTGTAAAAAGAGGGAAAAGCA AtmiPEP162a1 Arabidopsis CTAGAAGAAAAAACCAGATCTATAAAGTTTGTTATTAAAAGATAGAGAG SEQ ID NO: 220thaliana AGAGGAGGGATGTAGTAGGCCAATAGGCAAATCAGAGAATCACAAATGGTATCTGGTCAAGAAGATTCCTGGTTAAAACTTTCATCTCTCTGTTTCCTTTTTCTTTCTTTGTTGGATTCATTAATTTGACATATCTCTATCATCACACTGATTCTCTTTCTCCCAGTTTGTCTGCAGATGCATGTGTGTAATCTAGGGTATATGTTTTTGTCCATTTGGTTTCATAAGGCAATAAAGATCCAGCTATTTACTACTTGTGGTATAGATTTTGACTGTTGAATTTTCAGATCTGATGTGTTTCGTTTGATCCGATTCGGAAAATTTATGTTTCGTTGACATTTTGGAGTTTAGTTGGAAGAAGAGTGAGAGTCGCTGGAGGCAGCGGTTCATCGATCTCTTCCTGTGAACACATTAAAAATGTAAAAGCATGAATAGATCGATAAACCTCTGCATCCAGCGTTTGCCTCTTGTATCTTTCTTATTGACTT AtmiPEP162b1 Arabidopsis CTGCATCTATCCACCTCTCTCTGTAAATTTATCTAAATGTTTCTTTTAATC SEQ ID NO: 221thaliana TTTTTGAGATTAATAATGATTTGTGTTTGTTCATCAACCGATTTTCTCAGATCTGTCAATTATTTTTGTTTATTTATTTATGATTTATGAATGAGGAAAGAGTGAAGTCGCTGGAGGCAGCGGTTCATCGATCAATTCCTGTGAATATTTATTTTTGTTTACAAAAGCAAGAATCGATCGATAAACCTCTGCATCCAGCGCT GCTTGCTC AtmiPEP163-1Arabidopsis  TATCACAGTTCTCATCAAATATTTGAAAGTATCAAACAAAAAAAGGAGASEQ ID NO: 222 AtmiPEP163-2 thalianaGTGAGAAAAATAAAGAGAGAGATAGAGAGAGATCATGTCCACTACTCAAGAGCATAGGTCTTGATTGGTGGAAGACAAGTACCTTAGATAAACCGACCAAAACCCGGTGGATAAAATCGAGTTCCAACCTCTTCAACGACAACGATTTCAACACTCTCTTCCAGGAACAACTTCCTCCAGGCAGATGATACTAAAGTGCTGGAGTTCCCGGTTCCTGAGAGTGAGTCCATATCAAAATGCGCATTCGTTATCACTTGGTTGAACCCATTTGGGGATTTAAATTTGGAGGTGAAATGGAACGCGTAATTGATGACTCCTACGTGGAACCTCTTCTTAGGAAGAGCACGGTCGAAGAAGTAACTGCGCAGTGCTTAAATCGTAGATGCTAAAGTCGTTGAAGAGGACTTGGAACTTCGATATTATCCCCCGTGT AlmiPEP164a1 Arabidopsis AGTAGGGTTGGAAAATTTTTTTACATTTTTACTCTAAAATAGAATAGAGT SEQ ID NO: 223AlmiPEP164a2 lyrata TGGAGATGCCCTTAGCAGTTATTAGACAAGGGATTGTTTGGCCCTAGCGAAlmiPEP164a3 TCCTCTCTTCACTCTCTCACTTTTGTAGTTCAACCCTTCTTTTGCGTGAGATGCCATCATGGCATGACATGGTTCTTTTGCCTTACGTAAAACACACTCACGCCAACACACGCCACATAACATAAATAAATTATATATACATATACGTATGTGCGTGTGAGTCTTCCATTAATGCAATCTTTGGGCCTATATATATATACAAACCTTCCATAACCAAAGTTATCATACTACAAAAGCTCTCTCGTACTTGGAAATGCGGGTGAGAATCTCCATGTTGGAGAAGCAGGGCACGTGCAAACCAACAAACACGAAATCCGTCTCATGTGTTTTGCACGTACTCCCCTTCTCCAA CATGAGCTCCTGACCCATTGAtmiPEP164a1 Arabidopsis AGACAAGCCCCCACACTAAAAAAACAGTAATATGGAATAAAAAAAAGC SEQ ID NO: 224AtmiPEP164a2 thaliana TTTCAAAACTTAGCAGTTATTAGACAAGGTATTGTTTGGCCCTAGCTAGCAtmiPEP164a3 GATCGTTTAGCTCTCTTCACTCTCTCACTTTTTTAGTTCAACCCTTCTTTTGCGTGAGATGCCATCATGGCATGGTATGGTTCTTTTGCCTTACGTAAAACACACTCACGCCAGCACACACACACACACACATAACATATACGGATGTGCGTGTGAGCTAGTCTTCCATTAATGCAATCTTTGGGCCTATATATACAAACCTTTCCATAACCAAAGTTCTCATACTACAAACGCCCCTCATGTGCTTGGAAATGCGGGTGAGAATCTCCATGTTGGAGAAGCAGGGCACGTGCAAACCAACAAACACGAAATCCGTCTCATTTGCTTATTTGCACGTACTTAACTTCTCCA ACATGAGCTCTTCACCCBrmiPEP164a1 Brassica rapaAGACAACCCCACGTTTTAAAATAAGAAATGATGATAATTTTGTGGAAAT SEQ ID NO: 225BrmiPEP164a2 AAAAGCTAGTATACTTTTGCAATAATTAGACAAGGTATTGATGCTTTGGGBrmiPEP164a3 CCAAGCTAGTTTCTTTTAGCACTCTTCACTCACTAGTTTTTCTTCTCAGCCCTTCTTTTGCGTGAAATGCCATCATGGCATGGCATTGTCATTTTGCCTTTCGTAAAACACACTCACGCCAACATACATTATTCATATTCATGTGTATGTATATGAATGTTCCATTAATGCAATCTTTGGGGCCTATATATACGAAGCTTACATCACCAAAGCTCTCATATTACAAAAGCTCACATATATACTTGGAAATGTAGGTGAGAACCTCCATGTTGGAGAAGCAGGGCACGTGCAAACCAAAAAACATGAAATCTGTTTCATATGCTTTGCACGTGCTCCCCTCCTCCAACATG A CpmiPEP164a1Carica papaya AGACAACACTCCTCTTTGTTCCCTTCCTCACGTATCCACTTTTGAAATTTGSEQ ID NO: 226 CpmiPEP164a2TAATTTGTGTGCACCACCATGATTGCATGCCATCCCTACTTGCCTTTTCCCCTTTTCCTTTCTCTAACATTTTACTCAATCTTCTTCTCCCCCTCCCCCCCTTCCCCCTCTCTGCCATTATAACCATAATTAAACCTCTCTCCCTCTCTCTCCCTCTCTCTCTCTCTCTCTCTGGGTTCTCAGTATAAATGCAGCTCTGCTTATACTTCCACACCTATATATATATACCTGACCCTTCTTCACCTCCTTCATCCACCTCCTCCTTCTTCCCCAAAACTTTCTTAACTGTTCTCTGCATACATATATATCCACATACATATATATATATATAGAGAGAGAGTGAGACAGAGAGGTTACCGAGGCAATTGGGTGAGTAGCTCCCTGTTGGAGAAGCAGGGCACGTGCAAATTCTCCATGGCTTTCCCCTCTTTGCACGTGCTCCCCTTCTCCAACATG GGTTCC CrmiPEP164a1Capsella  CGGCCACCCCCACATTTAACAAGAAAAAAACTGATGGAATTAAAAGGTTSEQ ID NO: 227 CrmiPEP164a2 rubellaTGAGAACTTGGCAGTTATTAGACAAGGTATAGTTTGGCCCTAGCTTCTTT CrmiPEP164a3TAATTTAGCTCTCTCCACTCTCACACTTTTCAACTTTCACCCTTCTCTTGCGTGAGTCGCGAGATGCCATCATGGCATGGCATGGCATGTTTCTATTGCCTTACGTAAAACACACTCACGCCAACACATACTCACTATACATGTAAATAAGTATGTGCGCGTGTGAGTCTTCCATCCATCAATGCAATCTTTGGGGCTATATATATACAAACCTTTTCCATAACCAAAGCTCTCATATAAACTACAAAAGGCTCACTTGGGAAATGCGGGTGAGAATCTCCACGTTGGAGAAGCAGGGCACGTGCAAACCAACAAACACGAAACCCTCCTCATGTGCTTTGCACGTACT CCCCTTCTCCAACATGGrmiPEP164a1 Gossypium GAAAACCCAAGTTCAGGCTAACAAGTTATCTGATGATGAGATCAAGAAT SEQ ID NO: 228GrmiPEP164a2 raimondiiTTTAAAGTTTCAATATAGATTTGGCATGGGTATTGGCGGCAGAAAGCAAT GrmiPEP164a3TAAAAAACCAGTTATGTCAAATTCAAGGTCGTATCAGTTAAAATGAATGAAGATTTAGAAATTTCAACAAGGAAGAGGACCCCACAGCTTTGTTAAATTAAGTGTGTGGTTTTTATAATTATCATCTCGAAAGTTTCATAATATCAATTAGATTAAAACATCTCTGAATTTCATAATTACAAACCAGATAGATAGATACATGAAAACTTAGACCCCAGAGATCTGTCTTTAAAGAATGCCCACTTCTAGACTCAATCTCTATTACTCTCTTTTTTTCTCTCTCTCTCTCTTCGGAAAAACTTGTATATAAATAAATGACACTTTCTTTGCTTTCTGCACTCAACTCATGAACTTGAAAAGCTTTACTTGGATGGGTTGGTTGGGGGTGAGTATCTCTTGTTGGAGAAGCAGGGCACGTGCAAGTTCCTATGTTTAAGTGAACTTTGCACGT GCTCCCCTTCTCCACCGTGAGMtmiPEP164a1 Medicago GAAGAGAAAAAACCTAGTGTAAAATTTGATATACTCTTTATGTATAGTAC SEQ ID NO: 229MtmiPEP164a2 truncatulaGAATGTTTTTTTAAAAATTATGTAAAAAATGATAAAATAATAACTAACTAAATTAACAGTAAAATTAGAAAAGTAAAATACTATGCCCAAATTTGATATTTTTTTTTATATATTTGTATAGATTATTATTATTTGATATGTAAAGTCCAATTAAAAATTTGTTTTAACTAAGATTTGAACTAGGTTTTCTTAAAAGACTCATCTTTTACTTCAAATTTATTTATCATTTGAATTCAATCACTTTCTAATATTATTATTATTATTTCCACCATACTCATTGCTTCTGCCACGTTACTTTAGTTAGATCTCTTATGTCATATATCTCTCTCTCTCCTAAGTTGCTACCTATAAATACTAAGCCTTTCCCTTGGTTGGTTCAATTCAACTTCTACTTCTCATCAAACACAAAGTGCAATAAGCTTCATTTCCTGGGTGAGAAGCTCCTTGTTGGAGAAGCAGGGCACGTGCAAATCCTCTTTCTGATTCATTCTCTCATAATGCATATCAATATCTTTTGCACGTGCTCCCCTTCTCCAACTAGG OsmiPEP164a1 Oryza sativaATGCAAACCCACTCCAACACTCCACAATCCACATACTCTCTCTCTCTCTCT SEQ ID NO: 230OsmiPEP164a2 CTCTCTGAGTAGGAGTACATGTGTGTGTGTGATATCAATATGCATTCGATGTTGATGCTACTGTAGCCATCTTGTGGCTATATAAACCCAGCAGGCAGCAGCACAGCTTAGCTAGAGAGCCATATTGCATGCACACTCGCTAATCTCTTTTCTCTACTCTACTTGCATTACACCACCTCTGCATTGCACTTCAGTTCATTCATTCCACTGATGCATGGATCGATGTTGCTACCTTCTTCTCTTCTCCTCATGCATCCATGCATCGATCTCACCTAGCTTCTTCCTCATCCTCTCTCGATCGATTACAAGAGAAAAGTGTTTGCTGTTCTTGCTATCGATCTACAGGTGAGTAGGTTCTTGTTGGAGAAGCAGGGTACGTGCAAAATGCACACCGGTTGGTCGAGCTAATTAACAAGCTCTGACGACCATGGTGATCGAATGCACGTGCTCCC CTTCTCCACCATGGCCTAlmiPEP165a Arabidopsis CTAGGGTTTAGGAATGACGACTTGTTTCTGTTGTGTCTTATTAAAAGCCC SEQ ID NO: 231 lyrataATCTTCGTCTCCGCCACTCATCATTCCCTCATCATAACACCATCATCACCATTCACCAACCTCTCTCTCTTTCTCTCTCTCCTCTCGATCTACAACAAAATGTGAATCTGCTAAGATCGATTATCATGAGAATTAAGCTATTTCAGTTGAGGGGAATGTTGTCTGGATCGAGGATATTATACATATATACATGTGTATGTTGATACATGTGATCATAGAGAGTATCCTCGGACCAGGCTTCATCCCCCCCAACATGTTATTGCCTCTGATCACCATATATATGTCGTTACATTTCATGGTTAATTACTTGCACAAATCACAAAAGCTTGGTTTGTAACTTTCTATGACCTTTTTTAATGACTTTGAATCTTTCATGCATGACTTCTTAAGAGTAGATTTACACATTTGCGGATCCGTTTATGCTTTTTGCTTTTGTTTCGTTTATATATAT AtmiPEP165a Arabidopsis CTAGGGTTTAGGAATGACGACCTGTTTCTGTTGTGTCTTATTAAAAGCCC SEQ ID NO: 232thaliana ATCTTCGTCTCCGCCACTCATCATTCCCTCATCATAACACCATCATCACCATTCACCAACCTCTCTCTCTCTCTCCTCTATCACTCTCTACAACAAAAATTTGTGAATCTGCTAAGATCGATTATCATGAGGGTTAAGCTATTTCAGTTGAGGGGAATGTTGTCTGGATCGAGGATATTATAGATATATACATGTGTATGTTAATGATTCAAGTGATCATAGAGAGTATCCTCGGACCAGGCTTCATCCCCCCCAACATGTTATTGCCTCTGATCACCATTTATTGTTACATTTTTTTTTGTTAATTACTTGCGCAAATTACAAAAGCTTGGTTTTTGTGATGACTTTGAATCTTTCTTGCATGGCTTCTTAAGAGTAGATTTACGGATCCGTCTATGCTTTTTGCTTTTTGTTTCGTTTATTTGTATTTAAAC BcmiPEP165a Brassica GAGATCAATGAAATTATCCTGCCAAATAAAACGTGTGACGTTTATTCAAA SEQ ID NO: 233carinata AATATATGCATTAGATGCTTTGATATTAAAATATTTCCTTTTAAAAGCTAGCTAGGGTTTAGGAATGACGAGTTGTGTCTTATTAAAAGCCCTTCTTCTCCTCCGCCACTCATCATTCCCTCATCATAACACCATCATCACCATTCACCCACCTCTCCTCTTTCTCTCTCTCTCTCTCTCTCTCTCTCTCTAGAACAACAAGTGAGAATCTGCTAAAATATTGTGACTATTATCATGAGAATGAAGCTATTTCAGTTGAGGGGAATGTTGTCTGGATCGAGGATATTATATATACACAAATACGTATATATGTTAATACAAGTGTTTGATCATATATGTATATAGATTATTCTCGGACCAGGCTTCATCCCCCCTAACATGTTATTGCCTCTGATCACCAGATTCTATCAACTCTTCGCTTATTATTTTGTCACAAACAAGTAATAAGCTCATAATTTTCTTTGAGTCTTTCAGCATCGTTTCATTATGTTTTTCGAATCCG BjmiPEP165a Brassica GAGATCAATGAAATTATCCTGCCAAATAAAACGTGTGACGTTTATTCAAA SEQ ID NO: 234 junceaAATATATGCATTAAATGCTTTGATATTAAAATATTTCCTTTTAAAAGCTAGCTAGGGTTTAGGAATGACGAGTTGTGTCTTATTAAAAGCCCTTCTTCTCCTCCGCCACTCATCATTCCCTCATCATAACACCATCATCACCATTCATCCACCTCTTCTCTCCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTAGAACAACAAGTGAGAATCTGCTAAAATATTGTGATTATTATCATGAGAATGAAGCTATTTCAGTTGAGGGGAATGTTGTCTGGATCGAGGATATTATATATACACAAATATGTATATATATGTTAATATCAGTGTTTGATCATATATATGTATATAGATTATTCTCGGACCAGGCTTCATCCCCCCTAACATGTTATTGCCTCTGATCACCAGATTCTATCAACTCTTAGCTTATTATTTGTCACAAACAAGTAATAAGCTCAATAATGTCTTTGAGTCTTTCAGCATCGTTTCATATGTT TTCGAATCCGBnmiPEP165a Brassica napusGATATCAATGAAATTATCCTGCCAAATAAAACGTGTGACGTTTATTCAAA SEQ ID NO: 235AATATATGCTTTAAATGCTTTCATATTAAAATATTTCCTTTTAAAAGCTAGCTAGGGTTTAGGAATGACGAGTTGTGTCTTATTAAAAGCCCTTCTTCTCCTCCGCCACTCATCATTCCCTCATCATAACACCATCATCACCATTCACCCACCTCTCCTCTTTCTCTCTCTCTCTCTCTCTCTCTCTCTCTAGAACAACAAGTGAGAATCTGCTAAAATATTGTGACTATTATCATGAGAATGAAGCTATTTCAGTTGAGGGGAATGTTGTCTGGATCGAGGATATTATATATACACAAATACGTATATATGATAATACAAGTGTTTGATCATATATGTATATAGATTATTCTCGGACCAGGCTTCATCCCCCCTAACATGTTATTGCCTCTGATCACCAGATTCTATCAACTCTTCGCTTATTATTTGTCACAAACAAGTAATAAGCTCAATAATGTCTTTGAGTCTTTCAGCATCGTTTCATATGTTTTCGAATCCG BomiPEP 165a Brassica GGGATCAATGAAAATTATCCTGCCAAATAAAAACGTGTGACGTTTATCC SEQ ID NO: 236oleracea AAAAATATATGCATTAAATGCTGTGATATGAAGTATTTCCTTTAAAAGCTAGCTAGGGTTTAGGAATTACGAGTTGTGTTTTATTAAAAGCCCTTCTTCTCCTCCGCCACTCATCATTCCCTCATCATAACACCATCATCACCATTCACCCACCTCTCCTCTTTCTCTCTCTCTCTCTCTCTCTCTAGAACAACAAGTGAGAATCTGCTAAAATATTGTGACTATTATCATGAGAATGAAGCTATTTCAGTTGAGGGGAATGTTGTCTGGATCGAGGATATTATATATACACAAGTACGTATATATGTTAATACAAGTGTTTGATCATATATGTATATAGATTATTCTCGGACCAGGCTTCATCCCCCCTAACATGTTATTGCCTCTGATCACCAGATTCTATCAACTCTTCGCTTATTATTTGTCACAAACAAGTAGTAAGCTCAATAATGTCTTTGAGCCTTTCAGCATCGTTTCATATGTTTTCGAATCCG BrmiPEP 165a Brassica rapaGAGATCAATGAAATTATCCTGCCAAATAAAACGTGTGACGTTTATTCAAA SEQ ID NO: 237AATATATGCATTAAATGCTTTGATATTAAAATATTTCCTTTTAAAAGCTAGCTAGGGTTTAGGAATGACGAGTTGTGTCTTATTAAAAGCCCTTCTTCTCCTCCGCCACTCATCATTCCCTCATCATAACACCATCATCACCATTCATCCACCTCTTCTCTCCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTAGAACAACAAGTGAGAATCTGCTAAAATATTGTGATTATTATCATGAGAATGAAGCTATTTCAGTTGAGGGGAATGTTGTCTGGATCGAGGATATTATATATACACAAATATGTATATATATGTTAATATCAGTGTTTGATCATATATATGTATATAGATTATTCTCGGACCAGGCTTCATCCCCCCTAACATGTTATTGCCTCTGATCACCAGATTCTATCAACTCTTAGCTTATTATTTGTCACAAACAAGTAATAAGCTCAATAATGTCTTTGAGTCTTTCAGCATCGTTTCATATGTT TTCGAATCCGAtmiPEP 166a Arabidopsis CATCATCACCACTCACTTATCTTCTTCTCCATCTCTCTCTCTGCTTCTCCCT SEQ ID NO: 238thaliana TAATCTTAGCCGGGTCTCGTGGGGGACGAACATAGAAAGAGAGAGATATAAAGATATATATTCAGAAACCCTAGATTCTATAATTTCGACTGAAAAGAAAAAGGGGCTTTCTCTTTTGAGGGGACTGTTGTCTGGCTCGAGGACTCTGGCTCGCTCTATTCATGTTGGATCTCTTTCGATCTAACAATCGAATTGAACCTTCAGATTTCAGATTTGATTAGGGTTTTAGCGTCTTCGGACCAGGCTTCATTCCCCCCAATTGTTGCTCCCTGTTTACTCCATATTTCTTCCTTCTTTTCAAATTAGGGTTTCAGATCCAGTGAATGAACCCTTGTTAAAGGTTTGATCTCTT ACCTTACTTTAtmiPEP166b Arabidopsis TCTCATCATTCTCTTCATCATCACCACATTCATCTCTCTCTCTCTCTCTCTC SEQ ID NO: 239thaliana TCTCTCTCTTTCTCTTCCTTGATCTTAGCCGGATCTGTTGGGGGACGAACACATGAGAGATAGATAAAATATAAGAAATTTCTCGAAAAAACCTAATAGAAAAAGGTCTGTTTCTTAAAGAAGAAGAAGAAGAGGATTTAAAGAGGGATTTTTCTTTTGAGGGGACTGTTGTCTGGCTCGAGGACTCTTATTCTAATACAATCTCATTTGAATACATTCAGATCTGATGATTGATTAGGGTTTTAGTGTCGTCGGACCAGGCTTCATTCCCCCCAA AtmiPEP167a Arabidopsis TGATGAACAGAAAAATCTCTCTTTCTCTTTCTTGATCTGCTACGGTGAAG SEQ ID NO: 240thaliana TCTATGGTGCACCGGCATCTGATGAAGCTGCCAGCATGATCTAATTAGCTTTCTTTATCCTTTGTTGTGTTTCATGACGATGGTTAAGAGATCAGTCTCGATTAGATCATGTTCGCAGTTTCACCCGTTGACTGTCGCACCC AtmiPEP167b1 Arabidopsis AACCACAAAGTACCGCTGCTATTTTCTTTTTACGTCTTTGTATTTGCATCG SEQ ID NO: 241AtmiPEP167b2 thaliana TCTAAGAGAATGATGGGTTGTTTTGTGGGATTTTAATGCAGGAGGAAACATATGAGGGGTGATTAAGGCAAAAACCTTAAGATGTGGTCATTTAGATACATGGAGTCAAACTAAGAATGGACCTTGGCGAAAGCTTCTTCACGGTCAAGATTTAAAATCAGGTACGACACTGTGTACGTGAGAGAGAGAGAGAGAGAGAGAAAGAGATTATAGAAAGAAAGAGATGTATCACAATAAAGGAGTATATTTAGGGTCACAGGTGGTGGAGATATAGGTATGCAGGGCCAAGGCTCTAATCTCTTCATAGCCCTATTGATTTTGTCCCTCTCTCTCTCTCTTTCTTCCTCTCTTAGCTGTATGCATTATGATGCGTCTTTTAATTCACTGTTTCAGGCTTCTTTAATTCGTGGTGTCTCTCTCCTTTTTACCCAACCATCTCTTAAAATTTTTAACATCTGTTCCTCAAATCCTCTCTCATCTCTTTCTATAAGTATCTATAGCGCCTCTTAAACCACAAAGCATCACCTCTGTCTTCTCTCATCTCCTTTCTGTATTCTCTTTCATTGCCTTCACGTCTGTTGCAATTTCTCCACTTCTTGAGCTTCCGTTTTTTACAATTATTGATCCGTCAAATATGTGAGATTTGCACAACTTGTTGCTCAGGTATTTTGAAGACAAGTCCACAAGGGAACAAGTGAAGCTGCCAGCATGATCTATCTTTGGTTAAGAGATGAATGTGGAAACATATTGCTTAAACCCAAGCTAGGTCATGCTCTGACAGCCTCACTCCTTCCT AtmiPEP169c1 Arabidopsis GAGCAAGACAATGCCACATACAAACTTGAAAGATCTCTTCATCTTTTCTC SEQ ID NO: 242AtmiPEP169c2 thalianaCAAATGTTTTTTTTTCGTTTGCTATTTATCTCCACAATTCTTGGAACAAAAACTACATTCACAAACGAGAGAATTTTCACAACACCTCTTTTGCTCTCATTTTTTTTTTTTCGTCCATTATGAGTATTAATTATGGTTAGGGAATCTTACAGAATGAAAATGAAGGTGTGAATGGATTGTCTCATCTAAAGCCTTGAATGTGGGAAAAAGGCCATTGTTGTTCAGCCAAGGATGACTTGCCGGTAGCTTGTATTATGATTACTCTATATTCGATTTATATTATGGAGATGATGGTTTATATATATTTACTTATCTACATAGTTTTAGTTGATTTTTTTTCGTACGTAATATAATACGAAAAAGTATTTACTTATTTATATATGTGTGTTGGGGCAAGAAGTGTAACCAAGCTAGCCCGGCAAGTCATCTATGGCTATGCAACTGTCTCTTCCTCTCATTCTAGGCTTACGATGACACGTAAAAAATCCCAAATATCACTAAT d Inventor NationAtmiPEP16911 Arabidopsis AGGCATGAGACATAAAGAGAGTTAAATATAATGAAGAAGAGAGGTCTA SEQ ID NO: 243 thalianaATATGGCGAAAAGAGTCATGTTTAATAGCCAAGGATGACTTGCCTGATCTTTTTCACCTCCATGATTCAATTTTAAGTTCGTGGATTTTGGATTATTATGCGTTTAAAAGGTATAATAATTTGAGATCATGTTGAATCTTGCGGGTTAGGTTTCAGGCAGTCTCTTTGGCTATCTTGACATGCTTTCTTCATC AtmiPEP171a1 Arabidopsis GAATTTTGATTTATGAACCTCCTCAAGAAGGAAAGACAGAGGAGGAGAC SEQ ID NO: 244thaliana AAAGAAGTATAGGTTCACATTGCATAGCCAGTTTAGTTTTGAAGGATGGATATATGAAAAAAATATGAAGAGAGAGAAGAGAGAAGAAGAGGAGGATTAAAGAGGGTGAGGCCAGCTTTTGTGCTTTGGTAGTAGATGAGGTTTAAATGCTCCATACCTTCCATTTCCTTCTCTCTTACCCTAATTTAATTCTTCCTCTCCTTTATAACTCCCCACAGACATTCTCACTTCTCCTCCTCACACTTCACATCAACACTTCTTTCTTGTTTTTTCATTTTACAATGTTTCCTTTGATATCCGCACTTTAAGCATGAGAGAGTCCCTTTGATATTGGCCTGGTTCACTCAGATCTTACCTGACCACACACGTAGATATACATTATTCTCTCTAGATTATCTGATTGAGCCGCGCCAATATCTCAGTACTCTCTCGTCTCTATTTTGGACTTTGTGGTCTTGTAGATCGATTTGTATGTGTGTGTTGAAATGGAGACAAGTACTTGTAACTTCTTTGTTGTTATATTGTTTACCTATAGGCTGATGTCATAAACTCTTTTGATCTTGTTTCTAACTTCCAGATTCTTGAAAAATCAAGTCGTGTGTGTGTCTCCATGGAAGCCTTTTCCATTTCTTCCTTTCCA AtmiPEP171b Arabidopsis ACTCATAAACTTTGCTACTGGCCGCATTTCTATTTTCTCCTTCGATTCTTC SEQ ID NO: 245thaliana TAATTCGTACTTTGGTTTCTGACGTCCCTAAAATTTTTAGACAGTAAGAGTTTCTCCAGGATCCGATGGTTCTCTCCGGTAAATTAACATTTTAGTGTCAATAGTCATTTATACATATTTTTATTTCACTTTTTGTTTTGTTTATTGGTTTTCTGGAGCTAAGTGGAGATTATAGTCGAACAAGAGTGGTTTTATGCAAGGTAACGCGAGATATTAGTGCGGTTCAATCAAATAGTCGTCCTCTTAACTCATGGAGAACGGTGTTGTTCGATTGAGCCGTGCCAATATCACGCGGTAAACCAAAAATGGCAAAGATAGTTATTATAACCTTAAAGGTATGTATCATTATCGTTTTATTGTTTCAATTTTGATTAATGGCTTTGATATTTCATTTTTTTTTT MtmiPEP171b1Medicago  ATTGGTCAAACATACATACAGTAGCACTAGCTGGTTTCATTATTCCACTASEQ ID NO: 246 MtmiPEP171b2 truncatulaTGCTTCTTCATAGGCTCTCCAAATTTTGCAAAATTGAAAGAGACATAGTATATATATCTTAGCAAGGAGAAATTCAGGATATTGAGGATGAAGATTGAAGAGTAATCAGTGATGAAGAAAGCAAGCAAGGTATTGGCGCGCCTCAATTTGAATACATGGCTATAAAAATGCATCATATCAGCCATGTAGTTTGATTGAGCCGCGTCAATATCTTGTTTCCATCTCCAAATTTACCAATCTCATCAAATCAAATTAACACCACAATCAAGGCTTTCATTTAATGCAGTCAAAATAGGTTGACCTTATCATCGAAGAAATTGTTTTCTCATTCCTATCGAAGTTGGACTTGCCGAAAATGCTCGAAAGCATGTGTTTTAGTTCGACAGGCGAAAAAGTTACCGAAGGACAATTTGGTTGTGGTTCGGATAAGATCAAGCAACGGATATTTTCAAGACACGTTCGAAATTCAAATCAAATGGATAAGTATCGTTAGTTTACTGCAGTTATAGTTTTAAATTCAAATCTAGGCAGTTGTTTCTATTTGTATAAATAGTAGTTTTTCCCTAGGGAAAAGGGGTCGCAATTCAATCATACAAAAAACTTACAATCAAATTATCCGCATGGAAGAGAGAAACGAGTCACAAGTTGCAATGTATGAACATGTGTACCAATTTACATTCAATCAGTACAATTTAAGT TCATTTCCATAAAAAAAAAZmmiPEP171b Zea mays ACCAGGGTTAGGTATCCATCCACACAGCAATGCATCTGCCTTCAACTCCCSEQ ID NO: 247 TCTCGCCCCCCACCCCAACACACATCTCTCTCTTTTCTAGGGAAGGAAATGACGAAGGGGACGACGACGGCATGCTTCGGCTAGCTCGTTGGTGCTAGGACAAGGGCGGAGGTATTGGCGCGCCTCAATCCGAAGGCGTGGCTGATAGATTGGCGCGGCAGCCATGTTCTTGGATTGAGCCGCGTCAATATCTCCCCTTGCCTGTCCCGTACCTAGCTAGCTTGCTTGCCTCACTGATCGATGTCGTCCCTATTTCATGGAGAAGCTGATGATTGATTATTCTCACAAGCAAGAACTGTCTGATCTGTTGCCTGCATCGATCAGGATCTATATGCTGGAGAGTTCACAAGAACATGGACAGAACTCGCTTCAACAACCGATCAATCGATTGATTAGGTATGTACCTACCTCATATGCCTCAGCTCTTCGTTATGGATTTCTTCAACCGAAGGGTCAGTAAGCTCTTGGTTCCATGCCACTGCGTGAACTAAGCGTTCACAAAATCCGTTCCMCGGCATGAACCAAGCACTCAAAATCGCATGCAGCATCTTTCGTTTCAAAAAAAAATTGACTTYTGAAAACAATAGATGAATCAGTTTCAAACATATATGATTATCCATTTTCTCAACCGGGAATTTATATCTCGTTGGGATGCAAAACCGTTCAGTAGTAAAACTACTCCACGAGTATAAACTGTTTCAGTTATTTTACTATTAATTAGTTACCCGTATGCTGTTATGGTTTCTATATATCTATAAGTAAACCTTACTTAAATAAGATAGTTATACAAAAAAAAAAA AAAAA AtmiPEP171c1Arabidopsis  CAAGAAAAAACATTGAAATAGCTCATGTTGTCTCTTTCTCATTTTCATATSEQ ID NO: 248 thalianaCTGCTAAAAAAAGAACCGTGTTTTCTAAACTGGTTTAACGGTAAGTACCTGTCTCTAGTAACTTACCTATCAATTTGTTCCAATCATTTACTTGCTTTGACTTATTTGGTTTCCTTTTGTTTTGTTTTTCTTTAATATGTGGATGGAGTTTGGTGTAATAAGCAACTGAAGAGTCGATGAGCGCACTATCGGACATCAAATACGAGATATTGGTGCGGTTCAATCAGAAAACCGTACTCTTTTGTTTTAAAGATCGGTTTATTTGATTGAGCCGTGCCAATATCACGCGTTTAAATAGTTTAAAGATTCTATGTTAGTTGATGTGATCAATCAAGGTATGAATCTATATCAATTCTCTTATGCATAGTTTTATATTTACAGAGATGAGGTATTATCAATGTCTATCGTCGAGGATCACGCTCTTACTTATGTTATATTTCTATATAATTTTATTAATTAGTTTTCTAAAAGAGAAGGACAATTTAAAATTATTTTAAAGAGTTTTTTTTAAGTAGTTTTGTTTTCATGTTTATCTTCTGCAGGCTCTGAAGTTAGGATAGTAACAAGAAAAAAGACAGAAAAAAAGAAGAAAATTCATATACA TTCGTGA MtmiPEP171eMedicago  GAATAAGTGAATATTATCGATATTTATATCATATATCAACTTTTCTTCTGTSEQ ID NO: 249 truncatulaGCTTGCTTGCAAATTTGCAATTAAGCTTTTTTGATCTTATGTAAGAGAATATTATTGATGATGGTGTTTGGGAAGCCGAAAAAAGCGATGTTGGTGAGGTTCAATCCGAAGACGGATTTACATGTATAGAGTTGTAAAATACGATCTCAGATTGAGCCGCGCCAATATCACTTT MtmiPEP171h Medicago CCACAAAACTATAACTAGCTAGAAGCTTTAATCGCCTTATTTATTATAAT SEQ ID NO: 250truncatula AATAATAATAAATATGGCTTCAGCTGCAAAAGTATACATGGCGTGATATTGATCCGGCTCATCTATATCTTCAAGTTCAATCATCCATATTCATATCAATTTCAGACGAGCCGAATCAATATCACTCTTGTTTGCTTCATTGCATATTAATTATATACTTCATTTATAAGTTATAGTTTGCCATATATATATTAGATTGATTCTGCAGAAGTAGACAGGAGTGGTGTTGTTTCTGCTCATCTTATTAAATAATGAATGAATGAATGACATTTGCTTACTTATAAGACGAGCCGAATCAATATC ACTCCAGTACACCTAtmiPEP172a1 Arabidopsis CTCTCTCTCTCTCTCTCTCTCATCTGTGTTCTAGATCTCACCAGGTCTTTCT SEQ ID NO: 251AtmiPEP172a3 thaliana CTGGTTAATATATGGCTTCCAAGATCTGGTAATATGTTATAAATACGTCATACTTAAGCTTTTTTCAAATCAAAAATAGAAATTTGTGGGTTTGTCTCGTTTTACTATTTTAGCAGTATATATTAAGAAGTTCAGATGTTATTCGATCATCTGTTTTTGCTTCCCCTCTGCCATCTTTATCTTTTAGGGTTTCAATTCTTTTTCACTTTTTTCTTCTGGTTTGGAGATGGTTAGGTTCCAACTAAGTATACGAGATTAAATTTGACATCTTAGTTACTTCAAAATTCCTTCAATCAAAACAAGTCATCTCGACTATTCCGCCATGTTTGTATATACATATTTATATATTATATATATGAAGGTACGAGTTTCTAGTGTCTATAAATTAAGAAGGTTAAGTACCATATAGATGATATTTGTTAAGTAGTAAGTCACTCAAAGTTTGAGTTTGGGTTTGAGTTTGAGTTTGAGTTTGAGTTTGAGAGACAAAAGATTACTACAAGAAGATTGTTAAACAAAAATGGAAGACTAATTTCCGGAGCCACGGTCGTTGTTGGCTGCTGTGGCATCATCAAGATTCACATCTGTTGATGGACGGTGGTGATTCACTCTCCACAAAGTTCTCTATGAAAATGAGAATCTTGATGATGCTG CATCGGC AtmiPEP172b1Arabidopsis  ACTTGCACCTCTCACTCCCTTTCTCTAACTAGTCTTGTGTGCACCCATTTASEQ ID NO: 252 thalianaTGTGTACGTACTATTATCTCATAAATAAATATTTTTAAAATTAGATGCATTTATTGATATGAAAAAGTTACAAGATTAGTTTGTTGTGTGTGAGACTTTGGATCGACAGATCGAAAAATTAACTAACCGGTCAGTATTGAATATCAACTATTATATGCTCCATGCATTCGCTTATAGTTTCACACAATTTGTTTTCTTCACGGTCTAAAATCAGAAGATTCCATATATTTTCTTATGACGTAAAAGGACCACTTATAAGTTGACACGTCAGCCCTTGGATTCGTGAGGTTTTTCTCTCTACTTCACCTATCTACTTTTCCTCATATCCCACTGCTTTTCTCCTTCTTGTTCTTGTTTTTCTCGTTTTTTTCTTCTTCTTCTCCAAGAAAATAGAGATCGAAAAGATTAGATCTATTTTGTGTAGCAAGAAATTATCATTTTCGTTTCTTCATTCATATATTGTTCTATTATGTTGTACAATAATAGATACTCGATCTCTTGTGCGTGCGTAAATTTTATACAAGTTGTCGGCGGATCCATGGAAGAAAGCTCATCTGTCGTTGTTTGTAGGCGCAGCACCATTAAGATTCACATGGAAATTGATAAATACCCTAAATTAGGGTTTTGATATGTATATGAGAATCTTGATGATGCTGC ATCAAC AtmiPEP172c1Arabidopsis  TCACCAAATAGGCTCTTCTTTATCGCTTCATATATATAAAAGTCTACATCTSEQ ID NO: 253 thalianaATCTCTTTCTAGGTCACTAGCTAGACTCTAGATTAAGGATTGAAATTAGGGTTTCATGTTTCCAGCAAAATGGTGCCGTCTTGAGTCTTGAAAAGATCCAAGACAAAACCAAATCACTACATACATCCCTATCATCAACCAGCTACTGTTCGCTGTTGGAGCATCATCAAGATTCACAAATCATCAAGTATTCGTGTAAATAAACCCATTTATGATTAGATTTTTGATGTATGTATGAGAATCTTGATGATGCTGCAGCTGCAATCAGTGGCT AtmiPEP172e1 Arabidopsis TGTCATATTGAGAACTCTTTAGCCTTTGGCTTCTGTTCCTGACACTTGTAT SEQ ID NO: 254AtmiPEP172e2 thaliana AGTGAAGTGGGCTTGTGTTATATAGATGGGATCTCTCTCTTTATTTAAAAAtmiPEP172e3 GTCAATTAGAGATCTTGATGCTACTTCTGTCCCTTTCCAAGTGATTTTACGTCGACCAACTAGCTTTTTTCATATGAGTGTATATATTCATGTACCTATCTCTCTCAATTGCTTCTCACCAAAATCATCTTGCTGATTCATTTGCTGTCTGAATCCTCTTGCTTTCCTCTTTGCTTTTTCATTTGTTGATTTAAACCATGGGAGTTCCCAACTTTAGACCTCGAAACCGATAAGGATCTTTCTCTGCGGTTGAAATAGCTAGGTTCTCGATGAATAGGCTAGCCTTTGGTGGATGTTATCAGCCAGTAGTCGCAGATGCAGCACCATTAAGATTCACAAGAGATGTGGTTCCCTTTGCTTTCGCCTCTCGATCCGCAGAAAAGGGTTCCTTATCGAGTGGGAATCTTGATGATGCTGCATCAGCAAATAC AcmiPEP319a1 ArabidopsisTTGTATCCATAGTGTATTTCCTCGCATCTACCATCCATTTTCTACGCCTCT SEQ ID NO: 255AcmiPEP319a2 cebennensisCTCTCTCTCTTTCTCCATCAAATCTTGTTTTGTTCAAACTCTCTCTCTCATCAATTCTCTCCATACAATACATGCATACATACATACATACCATCTCTAATATTTCATCAATCTTCTTTTGTTCCAAACGCTCTTTCTCTCCATTTACATACATACGAATCATTGTTGTCATAGATCCGTTTAGAATTGCTTTAACTTTTAGATGAGATCTAGGGTTTCTTTCTTTTTCTTCAAAATCATGCTTTTTCGCTTGCTAGGTTATAGATCCATGTAAGTTTAGAGTAGATGTACACACACACGCTCGGACACTTATTAAATACATGTTGATACACTTAATACTCGCTGTTTTGAATTGATGTTGTAGGAATATATAAATGTAGAGAGAGCTTCCTTGAGTCCATTCACAGGTCGGATATGATCCAATTAGCTTCCGACTCATTCATCCAAATACCGAGTCGCCAAAATTCAAATTAGACTCGTTAAATGAATGAATGATGCGGTAGACAAATTGGATCATTGATTCTCTTTGATTGGACTGAAGGGAGCTCCCTCTC TCTTCTGTATTCCAhmiPEP319a Arabidopsis TTGTATCCATAGTGTATTTCCTCGCATCTACCATCCATTTTCTACGCCTCA SEQ ID NO: 256halleri CTCTCTCTTTCTCCATCAAATCTTGTTTTGATCAAACTCTCTCTCTCTCTCTCTCATCAATTGTCTCCATACAATACATACATACCATCATCTTTCCCATCTCTAATATTTCATCAATCTTCTTTTGTTCAAACGCTCTTCCTCTCCATATACATATACATACATACGAATCACATTGGTGTCATAGATCCGTTTAGAATTGCTTTAACTTTTAGATGAGATCTAGGGTTTCTTTGTTTCTTTCGTTTTCTTCAAATTTTGCTGCATATTCTCCAAGATCATGATTTTTCGCTTGCTAGGTTATAGATCCATGCAAATATAGAGTAGATTTACACACACACACGCTCGGACACTTATTACATACATGTTGATACACTTAATACTCGCTGTTTTTAATTGATGTTGTAGGAATATATATATGTAGAGAGAGCTTCCTTGAGTCCATTCACAGGTCGTGATATGATCCAATTAGCTTCCGACTCATTCATCCAAATACCGAGTCGCCAAAATTCGAACTAGACTCGTTAAATGAATGAATGATGCGGTAGACAAATTGGATCATTGATTCTCTTTGATTGGACTGAAGGGAGCTCCCTCTCTCTTCTG TAT AlmiPEP319aArabidopsis  TTGTATCCATAGTGTATTTCCTCGCATCTACCATCCATTTTCTACGCCTCTSEQ ID NO: 257 lyrataCTCTTTCTCCATCAAATCTTGTTTTGTTCCAACTCTCTCTCTCATCAATTCATTCCATACAATACATGCATACATACATACCATCATCATCTTTTCCCATCTCTAATATTTCATCAGTCTTCTTTTGTTACAAACGCTCTTTCTCGCCATATACATACATAAGAATCATTGTTGTCATAGATCCGTTTAGAATTGCTTTAACTTTTAGATGAGATCTAGGGTTTCTTTCTTTTTCTTCAATTTTTGCTGCATATTCTTCAAAATCATGATTTTTCGCTTGCTAGGTTATAGATCCATGCAAATATAGAGTAGATGTACACACATTCACGCTCGGACACTTATTACATACATGTTGATACACTTAATACTCGCTGTTTTGAATGGATGTTGTAGGAATATATATGTAGAGAGAGCTTCCTTGAGTCCATTCACAGGTCGTGATATGATCCAATTAGCTTCCGACTCATTCATCCAAATACCGAGTCGCCAAAATTCGAACTAGACTCGTTAAATGAATGAATGATGCGGTAGACAAATTGGATCATTGATTCTCTTTGATTGGACTGAAGGGAGCTCCCTCT AtmiPEP319a1 Arabidopsis TTGTATCCGCAGTGTATTTCCTCGCATCTACCATCCCTTTTCTACGCCTCT SEQ ID NO: 258AtmiPEP319a2 thalianaCTCCCTCTCTCTCTTTCTCCATCAAATCTTGTTTTGTTCAAACTCTCTCTCTCTCATCTATTCTCTCCATACAATACATGAATATACATACATACCATCATCTTCTTTTCCCATCTCTAGTTTTTCATCAATCTTCTGATGTTCCAAACGCTCTATCTCTTCATATACATACATACGAATATATTATTGTTGTCATAGATCCATTTAGAATCACTTTAGCTTTTAGATGAGATCTAGGGTTTCTTTGTTTTCTTTCAAATTTTGTTGCATATTCTTCTAAATCATGGTTTTTCGCTTGCTAGGTTATAGATCCATGCAAATATGGAGTAGATGTACAAACACACGCTCGGACGCATATTACACATGTTCATACACTTAATACTCGCTGTTTTGAATTGATGTTTTAGGAATATATATGTAGAGAGAGCTTCCTTGAGTCCATTCACAGGTCGTGATATGATTCAATTAGCTTCCGACTCATTCATCCAAATACCGAGTCGCCAAAATTCAAACTAGACTCGTTAAATGAATGAATGATGCGGTAGACAAATTGGATCATTGATTCTCTTTGATTGGACTGAAGGGAGCTCCCTCT BrmiPEP319a Brassica rapaTTGTATCCATTGTGTATTTCCTTGCATCCATCAATAAATTTTATGTTACGC SEQ ID NO: 259CTCTCTATTATTTCTCTCTACATCACACTGTCTTATGTTTAAGCTCTACTTCTCAGCAATTCTCTCCACCCAATACATGCATACATACCATCATCGTATCGCTCTAATTTTTCTATCAATCTTGTATCCTTCCACAAATTATCTTATGTCTCCCATTTTAAATCCTACATAGATCCACACATACGAATTATTCTTGTCTGAAGATCCATCCATTTACGATTGCTTTAACTTTTACATGAGATCTAGGGCTTCTTTATTTTTCTTCAAATCTTGCTGCATATATTTCAAGATCATGCTTTTCGGCTTGCTAGGGTTCTAGATCCATGGATGTATAGCGTACATACATACACGCACTAATTCATACCTGTAGTTTGTACGGAGAACATCATAAAATATCACTGTTTGGAATTAATCGTGTAGGAAATATAGATAGGTAGGGAGCTTCCTTTAGTCCATTCACAGATCATGATATGATCCAATTAGCTTCCGACTCATTCATCCAAATACCGAGTCACGAAAATTTCAACTTAGACTCGTTAAATGAATGAATGATGCGGTAAACAAATTGGATCATTGATTCTCTCTGATTGGACTGAAGGGAGCTC CCTC CpmiPEP319aCarica papaya CTATATCCCTGTCAAATAGTACTTGGTTTTGTTTTAGCCACAAATCTTCTGSEQ ID NO: 260 GTCTTGCAAAGTTCCTTCTGATCTCTCCCCCTTTCTCATTTTTTCCTCCTCTTATATATGGATCATCAATTTGCTGTACACACACACACATTTACTGTAGTGATAATTAGCTAGCTAATTTGTTAGTATGTAAATTAGATCCCAAGTACCCTGTTATATTTTTTTAGGCTTATCCTATGCATACCTGATAGTACAAGAACTTAGTTTGTAATTAGGTACTTGGTAGTAGGGTTAGATTAATTACTGTCTTGAAAGAGAACTTATCCAACAAATAGAGCTATGAAGATTAAATTAGGTTTTAGTCTTATTAAGATTATTATATTACTAGACAAAAACAGTTAAATTTTTTTAATTGGGTAATTAGGTACTTAGCAATAGGGTTAGATTAATTACTGTTTTGAAAGAGAACCTACCTACAAATAGAGTTGAAATGATTATGTTTTAGTCTTACTAAGATTGTCATATTTCTGGAAAAAAACAAATCTTGAAACAGATAATTCAGATAGTCATGATCAATGGAAAAAACATCATGGGTGTGTGCTTAATTAAGCTAATATATATATATATGAAGATATAATGTTATGCACACTAGCTATGAATTTGTAAGAATAATGAAGGATAAAGATGATATATTTAGATGTTATAAGTGTAAGTAAGGTGGAATGGGTTGATGGGTAGTAGTAGTAGTAGTAGAGATGATTGGTGGAGAGAGCTTCCTTCAGCCCACTCATGGATGGGTATGAAGGGGTAGAAGTAGCTGCCGACTCATTCATTCAGCCACTCAGTATGTAAACTCGTCCCACTGTTGACTGTATGAATGATGCGGGAGATATTTTTACATCCATCTTTCTCTGTGCTTGGACTGAAGGGAGCTCCTTCTT CrmiPEP319a Capsella TTGTATCCATAGTGTATTTCCTCGCATCTACCATCTACTATTTTCTACGCC SEQ ID NO: 261rubella TCTCTCTCTTTATCCCTCTATCTCTTTCTTCATCAAATCTTCTTTTGTTCAAAGTCTCTCTCATCATTTTTCTCTATACACATACATGCATCCACATACATACATACATATACCATCATCTTCTTTTCTCATCTCTAGTTTTTGTTTATAAATTTTGTTCCAAGGATCTGTATCTCTCCAATAAAGATACATACAAATTATTGTTGTCATAGATCTATTAGAATTGCTTTAACTTTTATATGAGATCTAGGATTTCTTCCTTTTCTTTCAAAATTTGCTGCATATTTTTCAAAATCATGATTTATCGCTTGCTAGGTTCTAGATCCATGCAAATTTAGAGTATTTTTACACACACACACGCTTGGACACAAGTACATACATGTAGTTTTCTTTTATGTGGTGAAAAGTACATAACATGTAGTTTATAGTTACTAGTCGCTATATAATTTAAAATTGATGTTATAGGAATATATGTACGGAGAGAGCTTCCTTGAGTCCATTCACAGGTCGTGATATGATCCAATTAGCTTCCGACTCATTCATCCAAATACCGAGTCTCACCAAAATTGGAACAAGACTTGTTAAATGAATGAATGATGCGGTAGACAAATTGGATCATTGATTCTCTTTGATTGGACTGAAGGGAGCTCC EgmiPEP319a Eucalyptus TCGTTTCCCATCCCCATTTCATAGAATAATGCCACCAAACAAAGAAGCAT SEQ ID NO: 262grandis TAGCTCAAAGACTAATTACCATCTGTTTTATTGATAGATACGTGCGAAACGGTGATTGTTTTTTCCCAAATAAGAAACCAAAATGAAGCATATTCAAAGGTGGAGATATGGGGAGACTTCCGGAAGGCAAGGGGATTGGAAAAGGCTCGAGATCAAAGTGCATAGCAACCCTTCGCTAAAGGTGAAAAAGAATACGAATAACTTCAGTAGCTCACTTTAAATTCCGAAACATTAAACAAATCAAATCTCCCTCGCCCTCCTTGCCTCCTCTCTTTACCTATATAAAGCCACCGCCCCTTCAATGAAATCCACGAGTGGAAGGTCACAGTATAGTAGGGTCCTGCAAAGGGAGAGCGAGAGCGGCTCCACTGTCTACCTATAAGCAGTTCCTTTCTTTTGTTTACATGTCTGTTGCACCTCACCGAGTTTTTCTATTCTCTTTCCTCTGGTTGGTTAGCAGATTTCTCAGGGGACTTCCCTCCTCCCTTGAGGATCCTTCTCTTGAAGCGATATGTCTCGAATGGGTAAGAGAGAGAGAGGAAGGGAGCTTTCTTCAGTCCACCCATGGGACGTGTTGGGTTTTAATTAGCTGCCGACTCATTCATCCAAATACCGAGCGAGAGCAAGTAACAGAGCTCCGTAAATGAATGGATGATGCGGGAGTCTTGTTGATTCCCAAGCTTTCCGTGATTGGACTG AAGGGAGCTCCCTCTATCTGrmiPEP319a Gossypium GCGTATCCTTCCACTTACGGATTCACCATAGCTGCTATAGCCCGAGTTTG SEQ ID NO: 263raimondii CTTGTCATAATAGAAATAAACTAAGGGAGAAAAAAGCTCTCCACTCTCGCCTTTTTCTTCTTCGAGTCTCTCTCTTTGACTGGCCTTTGTGCAAAGATCTTCCTTTTTTAAACAGTCGCTTTCTTTACTCTCCCTTTCCCTTCTTTCTCTTAATTGCTAAAGCAGCTTCCACTTCCACTCACTTTAACCCATGATCCATTTCAACCTGTCACAGTGGAGAGCAATTTGTATGGCTAATTTCCATCTCACCTATTCTTTTCTGTTTGGGGTTCTCTAGTCTAGGGTTGCATGACATGAGAGACATGGCTCTTTTTTTTTTTTTTCAGTTCACAGGTGTTTATATATGTTGTTGTGTTATTTTGTCTTAAAGCTTTGTGATTGATGATCTGATAGGTAAGAGAGAGCTTTCTTCAGTCCACTCATGGGATGGGGATGGGGTTTAATTAGCTGCCGACTCATTCATCCAAATACTGTGTTACAAAACCCAGTAAATGAGTGAATGATGCGGGAGACAAATTGAATCCTAATCTTCCTGTGCTTGGACTGAAGGGAGCTC CCTCCC MtmiPEP319aMedicago  ATTTATCCAATCATGTTGCTCTCACTTCTTTTCAGCAGTTCTGTTGATATCSEQ ID NO: 264 truncatulaACCCTTTTAGTTGAAGCTTTGGATTCATGACTTTATGAGATGGTAAATCTTTAAATTAAGTTTAAGTATGCACTTGATTTGTTTATATAATTTGTTTATTTAGATTTTAAACCCTAAGTTGACTTTTTTAATTTGATTTAAATTATGATATGATTGTTATTTGGCTTACCATGGTCATAATTTAGGGTTTAGAAGATGCATGTATATCTTGAATTGTTTATGGTAATAAAGGGGTTAGGATTTCTCCTTTTGGTGAAGTGAGAAAATCTCATAATTTTGTTCTGAAGGTAGTTTTTAAGATTTAGGGTTATGGGTTCTTTGTTTGAATGCTTTTTCAAGTCTTTTTCAATGATATTTGCCTAGATCTGTTTTAATTTTGAATTAAAATTCTGGGTTTGGATTAATATTATTGAAGATTATTATTAATTAATTTATTAGAAATAGATGAAGAGAGCTTCCTTCAGTCCACTCATGGAAGGGTAAGGGGTTTGAATTTACCTGCTGACTCATTGATTCAAACACAATAGACAATTATGGGGTTATGCTATTGTGAATTGTGTGAATGATGCAGGAGGTGAATTTCTTCCTTTTCTTCTTTGCTTGGACT GAAGGGAGTCTCCCTTTOsmiPEP319a Oryza sativaTTATATCTGACGCGTTGTAATCCTGTTTAATTAGGGCTTTGCCCATTTCTT SEQ ID NO: 265TTGACCCCTTCCGGACATTCGCTAGTTGGAACCTTGTTTTACTCCTAGCAGTGTACTGTGTAGTACTTATTACGAGCAAACGTAAAAATAAATAAATGGAAATGATACAAAGGCCGTGTTTAATTTTAAAATTTTTTTTCAAACTTTCAACACTTTACATCCCATAAAAAGCTTACTATACATACAAACTTCCAACCTTTTCGTCACATCGTATCTAATTTCAACTAAACTTTTAATTTTAGCGTGAACTAAACACAGCCGAAGCCCGGCCACATTCTCACTATTTTTATTCATTTTATCATGCCTGTGATGTCACGCCTTGGCCCTATTTAATAGGCCTTCTCCATTTCTCTCCATATGATGTCTTCTCTTCTCTATCCCTCTTGCCATCTTCTATCTTCCCTCTTGCACCCATCTTTGTGATAACTTCTACTAGCTCCTCTCCTACTACCAGTCATACCACTCTCACAAATCCTCCAAGATCCGCATGGGGAGAAGCTCCAAAAGTTTCGTGGTTAGTTTAATTTCATGCTTGTTTGCTGCCGTTTTTCATGTTGATCTGATCTTAATATATGTAGACTGCTGTTAACATATTCTTTTAATTTGATGGAAGAAGCGATCGATGGATGGAAGAGAGCGATCCTTCAGTCCACTCATGGGCGGTGCTAGGGTCGAATTAGCTGCCGACTCATTCACCCACAATGCCAAGCAAGAAACGCTTGAGATAGCGAAGCTTAGCAGATGAGTGAATGAAGCGGGAGAGTAACGTTCCGATCTCGCGCCGTCTTTGCTTGGACTGAAGGG TGCTCCCTCCTPpmiPEP319a Physcomitrella TGGGATCCACACGAGTAATTATTCCCTCTCTCATGCATCACAACTTGGAC SEQ ID NO: 266 patensTTGCCCAGCTTTTACCTCTCTCCATGTTCCACCGTCGGAGATCCTCGGTGCTGCTACCCCCGTTCGGCCAAACCCAACCCAACCCTAGGTGTCTGCCGGACCTCCGCTTCCCCTCCTGCTTCACCCCCTGCACCGCTTAAATTTGCACCTCGTAGTATTTATCTATGCCCCATTTCAGATGACTTGCATGACACCCATCTGGATTCGTCTCCAGGCGCCTTGTCGTATCTATTCGCTATCAGTCTTTCTTTCAGTCTGTCTTTCTGCCAAGCACACCGTGCTGGTGGTGGTGCTATCAGTCGAAGCAGTCGCCGAAGGGTCCTTGTATCCAGCCCTCACCGGAGACAGTTCGCTTCGGGGTGGGGAGAACTGTTGAGACCGTCGATGTTGCATGCAGCAGCACTGGCGAAGGTGCTTCTGATACTGGGTATTCCAGCCTCGCCGCTCGGTGCACTGCAGGTACGTGGTTACATGACACTACTGTGTGGGTAGTCTGGGCACAAGTAGATCGACATGCCAGATTTGGCCCGTATGCGTGTATGTGCGGTTATGTTCCCGTTTCGATTGCGGATACCTTGATTGTGGAGCTCCGTTTCGGTCCAATAGTGGCTGCGACGGAAGGTGGTCCCGCTGCCGAATCACACGTCCGGGTTGCTTATCGGGGCAGGGCCCCGATACGGTATCCGAACGTTTGTCCCGGGAACTGGTCGACCTTCCGCCCGGCGTCTCTTGGACTGAAGGGAGCTCCACT ThmiPEP319a1Thellungiella TTTTATACAAATAATGTTCGATAACACTAAACCCTAGCCATCCAACTAATSEQ ID NO: 267 ThmiPEP319a2 halophilaAGACAAAACCCTACTTGTAATTTACAACCGCAAATTCCCAGAGAACAGAGTAACTACGAGAGAGAGATGGAGATTCAAATTAAAAAGAAAAACTTATATATAATGAATACACAAAAGCTACCTAATCTGTATATATATATATATAAATATGTCTTCATTAAATTAATGGTCGTGGAATAGAAAAAGGAAAACCTAATTTGATCGCTAGGGCTTATCAGAGTAAAGATGGTTAACCTTCAAAAGATGACTAATTAACCGGGGAGATAATTAAAAGATTAAATACGCCAACAGAGAGTTAAGAGATACCAGATTTAAATTCCACAATTTGGTCATGTTCTTCTTCACGTATTCATGACGATGTCTGAATTATAGAGAAACCCAAAATATAAAATGTTAATTTTACCAGACATTTACATACCAATAACTCTATGACGATATGTAAAGTAAGCAAGGCATGTTTTTATGCAGGGAAGATTGAAAATTCAAGATTAATCAAGAAAATTGGAATACCAAAAAGAGAGGGAGCTCCCTTCAGTCCAATCAGAGAGAATCAATGACCCAATTTGTCTACCGCATCATTCATTCATTTAACAAGTCTAGCTCGAATTCTTGGTGACTCGGTATTTGGATGAATGAGTCGGAAGCTAATTGGATCATATCACGACCTATGAATGGACTCAAGGAAGCTCTCTACAAATGTATTCCTACCACATCAACCCAAATATAGTGATTACAGATGCTGTTCTCACTGTAGACTACATTTACGTTT AtmiPEP319b1 Arabidopsis AGACATCTCTTCTTCTCTCATCTCTCTTTTCTTCTCTCTTTTCCTCACATAA SEQ ID NO: 268thaliana ACTCTCTTTTTTTACTATTAAATCCATATGGTACCTCAAATTAATCTATGGTCATCTAGGGTTATCTTGAAGATTAGAATTGATTCTAGCACGCACAGAGAGGAAGATCATTGCATCCAGAATCACAAACATGGCCTATCTTTTATCTTTTCTTTTTGATCTAAGTCACTGTTTTATGCTATATATAGTATAATCAAATTCTTTACATGTGCTTGTATGTATGCGTATATATAGTAACGGAATTGTTAATATGCTTATAGATGTTGAGTTGGTGGAGGAAGAGAGCTTTCTTCGGTCCACTCATGGAGTAATATGTGAGATTTAATTGACTCTCGACTCATTCATCCAAATACCAAATGAAAGAATTTGTTCTCATATGGTAAATGAATGAATGATGCGAGAGACAAATTGAGTCTTCACTTCTCTATGCTTGGACTGAAGGGAGCTCCCT AtmiPEP394a1Arabidopsis  TCTTATTCCATCACAATCATCTAGGGTTTTAAGCCAAGCTTATATAGCCCSEQ ID NO: 269 thalianaGTCATAAAGAGAACTCATCTGCCTCTCTCTCAATACCAATAAATATCACCACCGTCCTTCTCTCCTATCACTATTCAATCTATCGCAAACTCCTTTATGTCTCTCCAATTTTATGAGAGGGTTTCCTTCAAGAACACAGTAAAATAGATTGGATCTTTAAACTTTTGTTCCTTTTCATGAGGGTTTGACAAAGATTTTCTTACAGTCATCTTTGGCATTCTGTCCACCTCCTTCTATACATATATGCATGTGTATATATATATGCGTTTCGTGTGAAAGAAGGAGGTGGGTATACTGCCAAT AGAGATCTGTTAGAtmiPEP395c1 Arabidopsis TTGTATCATGACAGAGCAAGAAGAAGAAAGTCAAATGTCCACATGAGTT SEQ ID NO: 270thaliana CCCTTTAACGCTTCATTGTTGAATACTCAAAGCCACATTGGTTTGTATATAACACTGAAGTGTTTGGGGGGACTCTTGGTGTCAT AtmiPEP395e1 Arabidopsis TTTCAAACCCTAACACTCTTATAAACCGATTCGCCAAAATGTATCTACAA SEQ ID NO: 271thaliana TATATTGATAATGTAATATCTATATATTCAAACAATCGTCGTGTTGGTCGGATGTTTTCTAGAGTTCCTCTGAGCACTTCATTGGAGATACAATTTTTTATAAAATAGTTTTCTACTGAAGTGTTTGGGGGAACTCCCGGGCTGATTCGGTATTTTAAATTCAGTAGACTAGCTAGCTG AtmiPEP397b1 Arabidopsis TGGTAATAGAAATGAGCAAGGAGATATTTTTTTCCCCTGGGTTTGAATGA SEQ ID NO: 272thaliana ACATCATTGAGTGCATCGTTGATGTAATTTTACTTATTTTATTCCATTGTTGAATTAATTAAAGAAGTATATATCAGCGTTGCATTCAATTATGTTTTTCTAATTTTCAGGAAATACAAAAAAAATGAAAAAAAAAAATCACTTAAAAG ACCTTGAGAGTTCTTTTGACTAtmiPEP398c1 Arabidopsis GGATATCGAAACTCAAACTGTAACAGTCCTTTTATTACTGGTTTAGAAGA SEQ ID NO: 273thaliana TAGATAAATATTGTTAAGGTAGTGGATCTCGACAGGGTTGATATGAGAACACACGAGCAATCAACGGCTATAACGACGCTACGTCATTGTTACAGCTCTCGTTTCATGTGTTCTCAGGTCACCCCTGCTGAGCTCTTTCTCTACCGTCCATGTTTTATCAACGCCGTGGCCCGTG AtmiPEP399b Arabidopsis TCTTATAGAGATGAAGAGAAACATGTAAACTCACTAGTTTTAGGGCGCCT SEQ ID NO: 274thaliana CTCCATTGGCAGGTCCTTTACTTCCAAATATACACATACATATATGAATATCGAAAATTTCCGATGATCGATTTATAAATGACCTGCCAAAGGAGAGTTG CCCTGAAACTGGTTCAtmiPEP399d1 Arabidopsis CAATAACTCAAAATGCAATGTGAAATATGAAGAATATATTAAATAGTAG SEQ ID NO: 275thaliana TGAAGATGCATGTTTATGAAGACAGAGAGATAATGTATGGTTGGATTACTGGGCGAATACTCCTATGGCAGATCGCATTGGCTAGATATGCAAGTAAAATGCTTCTCTGCCAAAGGAGATTTGCCCCGCAATTCATCC AtmiPEP403 Arabidopsis ATTTAGGTCTCTCTTCTTCTTCTTCTTTTTCTTCTTGAGCGCCGGCGAAAA SEQ ID NO: 276thaliana AAGTCTCTGTGAGAAAAAGATACGACGATTGTCATTAGAAGAGTCGTATTACATGTTTTGTGCTTGAATCTAATTCAACAGGCTTTATGTAAGAGATTCTTTAACAATTCCTATAATCTTTGTTGTTGGATTAGATTCACGCACAAACTCGTAATCTGTCTTTTCGATTTTTACCAGATCTGTC AtmiPEP447a1 Arabidopsis AATTATATCCATGGTCATGGCTCATCATTAGTCGCACTGCTCTCCTTTTCT SEQ ID NO: 277AtmiPEP447a2 thaliana CAAAGTTTAAATTCGACATTTGGTAAAATGATGAAACCTCGATGGAACTGCTCTCTTTATGGAATCACGGAATGGACAAATAATCAAAATCAGAAATCGAAGCGAAAAGGGAGGAGAAAAACGCAGATTTGGAGGATTGGGGACAGATTAGATACTGTTGAATGCATCACTCTAATGCTATCAGCCTATTAATAGCGTCCTATATTTTCGAAGACTTTTAATGTTTAGGGTTATGGATTTTTCGAGCGAAGCATGGAGAGATGTTGAATTGGATACTATAGGATTTGGTACAACACATACATATGTTCTGCTTCTGCAAAACTAACATATCAAGTTCAGAGAAACCAGTAAGTCGTTGAATATTTTATTATCCATTCAACGCTTTCTTCTTTTGGATCATGTCTTGTTTGCTTGACCACTTCTTCTTGCTTAAGAGGATGGACAATATATAAAAACTGGAGCCTTCTTTTTCTATGAATGCTTATCATCGCGGAGTTGATCTGTTCAATTCACCTGCCATTGGATGCTTTTTTTATATATACTTCACTGTTCAATTTCAGATGCTTTAGAAGGTTTGCGGAGTAGCTAGAGAATCTGGTATCTTCAGTTCTTCAATTTCAGCTACTTGGTATCAGCTTCGTCATTGTATATCAACACATTCTTAATATATAATACTACTTTTTCATCCATTAAACCCCTTACAATGTCGAGTAAACGAAGCATCTGTCCCCTGGTATTGTCTTCGAGCTTGGTGTTTTTTTCTAGCCAACTCCAAGTTCTCGAGTTGATCATTGTTTGTATTCTTGAGACATTATTTGGGGACGAGATGTTTTGTTGACTCGATATAAGAAGGGGCTTTATGGAAGAAATTGTAGTATTATATATCGAGAGTG AtmiPEP447b1 Arabidopsis CTATAAATGCTGCTTATCATCGTGGAGTTGGTTCTGTAAACATTTGAAAA SEQ ID NO: 278AtmiPEP447b2 thalianaTTCTGAACAGTTTCACCTGCCATTGGATGCTTTGTTTCAATTTCAGGTGCGTTAGAAGGTTTGCAGAGTAGCTAGAGAATCTCGTATCTTCACTTTCTGCTACTTGGTATCAGCTTCGTCACTTTATATCAACACATTCTTAATATACAATACTACTTTTTCATCCATTAATCCCCTTACAATGTCGAGTAAACGAAGCATCTGTCCCCTGGTATTGTCTTCGAGCTTGGTGTGTTTTTCTAGCCAGCCCCAAGTTCTCGAGTTGATCATTGTTTGTATTCTGACACATTATTTGGGGACGAGATGTTTTGTTGACTCGATATAAGAAGGGGCTTTATGGAAGAAATTGTAGT ATTATATATTGAGAATGAtmiPEP447c Arabidopsis TAGTATAACCGCTGATGTACACCTACCAGCTTGATAACTCTTTTTCGTGG SEQ ID NO: 279thaliana TTTCTGTGTACTCGTTTCTGTTTGTACAGATACTTCTTGTTCAATTTCAGATGCTTTAGAAGGTTTTCGGAGTGGCTAGAGATCTGTTATCTGTATGAACAGCTACTTGGTATCAGCTTCGTCATTTTATCAACACATTCTTAATATACAATACTTCTTTTTCATGCATTAAGCCCCTTACAATGTCGAGTAAACAAAGCATGTGTCCGCTAATATTGTCTTCGAGCTTGGTATTTTTGTATTCTGATACGGTATTTGGGGACGACATCTTTTGTTGACTCGATATAAGAAGGGGGTTTGTGGAAGAAATTGTAGTATTATATATCAAGAATG DmmiPEP1a DrosophilaTTTGTGGAACACATTCGACCCACTGAAAAATTGATATAATTTAATGAAAG SEQ ID NO: 280DmmiPEP1b melanogaster TGCATAAAAATGGTGGACAGTGCATTAAACTGAGCATTGAACACAAAGGCCGCTCAGCAAATTGCTAATTAAAATTCACGATTGCCATTTCACCTGACACGTTGACGATTTTCATTACAATTCGATTATGTTTCGTTGCAGGGAATTTTAAATGTTAATTGCCAAGAATGTTTCAACAAATTCATTTCTCATTAATGTGTCTTTTCATTTAATTTTATGTTGTATGAGCTGCACGAGAAATGAGTTGTACTTTTAGTTCGACGGCAGAGTCATGAATGTTCGGCAAAGAATGTAATAATAACTATCCTCTTTAGACAAATATAGATACAAATCTATCAGATTCTAAAAGTAGAATAATCAATTAATCAGAAAGCTAAAAATAAATAGGCATATTTATATTTTAATGCGGATTTTTGAAGTTCAACGGGAGAAATGAATCCTTTTTACCAGCCACAGGCGCAATTTGCAACAGAAAGTGTAGCAGAAGTACTCCTCGAATATTTCCCTGCTCCAGGAGTCATCCATGTGGTTTCGAGGCACACATTTGACAAACTCATGCCCCGCTATTTGTTGTAAAAACACAATCGCACACATGGCCGCATTTCGGCGACTTCCAGAGAGCGGTACACTTAAGGCGGCCTGGGAAACGCCTGCAATCTGCTGGTCGCGAACTGCAGATTGCATCCATGTGCCAGGCGACCATGCGACCATGTGACCATGTGCCCGCCCGACGCCTCGCAGCCCACATCCTGCCCATCGAGGGCACAACTCAGCGTGGGTATTGCCGCTCCGGCTGCTTCAAGTAGGTAAAAACCGAGAAGATTGAGGATGAATGTATGAGTATGAGAAAATACTCGGCGGAACATATGCTGCCGGGCTTGACCTGACCCTGCCTCATGTGTGGGTCTCCGATTTAATTTTAGGCACCTATATAAACGCGTGTTTACACTGCAGCCAGAACACAGTCGCCGTCTTCAGTTCGCGCCGTCAACTCCTCGATCGATCGATCGCATCGTCTCGGATCGAATAGAGCTGGGCTTCTGCTCCGGAGCTACATCGCCGTACTTGTCGGACGAGTGTGGTGATGAAAAGTCGCTTAGTCCGGGATTCCTGCCAGATCTCTAAGGGATGAGCTGGCATCCCAGGCTGGCCATGTGGCGCGAAGTATGCGCACAAAAAAGTCAAACAAAAAGGCGCAATTTTATTACGGGCAACCAACGACGAAACAAAACAAAAGCCAACCGAAAAGCAGAAACAAAGCAGCAAAAAGTTTATGAATTTTTTGTGCAGGCGCGTGAAAGATGCAAAACGAGAAAAAAACATGAAAAAAAAACATTAAAAAAAACAAAAAAAATCCAAAACAGATACCGAGCTGTATCCGAAAACGAGTGGGGAAAGGGGTTTCCCAGTCACATATAAACACACTTCAGTGCGCTTAAAAATTGCTTTATTGCAGTTGGACTATAAAAACGCACGGCAGCGAACACCGCACAACAAAAAGGACGAGCAGAAGTGGGCAAATAAAACGAAAGCTCTTAAACGAAAAACAGGAAAATTTGCATGCCACAAAAATAAGCATAAGGATTTGCCGCGCACAAAGTAGAAGCAAAAAGGAATTGCCCAAATGCAGCCACAAAAGACTGTGGCAAATGTTTTGCAGCTTGCCCCTTTTTCCCTGCAATTACCGTCAGTCGTTGTCATTATTCAGCAGATTATATGGTTTTGCTTATTCCGGACCACCATCATCATCATCATTATCATCATCTTCGGTAAGTTAGACAATCCCATAAAAAACTGTCCAAGTGAGTAGTGCCACCAAAAGTTAGCCGCGTTGTGGAAAATCCAAAACAAAGACCATCCCATATTCAGCCTTTGAGAGTTCCATGCTTCCTTGCATTCAATAGTTATATTCAAGCATATGGAATGTAAAGAAGTATGGAGCGAAATCTGGCGAGACATCGGAGTTGAAACTAAAACTGAAATTTGATTGAAACAGAAGTAGAACCGTAATGAAATGAATGAAATATTAACCCGTTTCTACAATCCCTGAATAAAATTATTAATTAATTATAGAGCGGGCTAATTTTACAATATATATTGATTTTTTTTTGAAG DmmiPEP8 DrosophilaATTCTTTTTTGGTGCTCGATCGTGACGGTTTGCTCGCGCTCTCCGCTGCGC SEQ ID NO: 281melanogaster CGCTCTTTCCGTTGCATATGTGTGCGGGCGTTATTGTGCATGTTTCCGGTGGCCGAAAAAAAATAGTAAAATAAAATATAGAAAACAGAAACCAAGAATAATAACAGCCATACGATAAACAGTGTGCCAATGTGTGTGTCTGTGTGTGTGTGCATCTCGCGTAACAACAATAATTGCATTTATCGGATGGCGCCAGCTTCAATTTAATTATAAATAACATGTTCAACTTTTTATACTATTTTCCCTGCGTCAAAGTGGGCGTTGCAACTGCCCCCGGAAAATCACGCGCCCCGGTTCAAAGTTAAAGTTTGCTGGGTAACGCACACACACACACACACAATCACTCACACGCGGTCACACGCACATTTCAATAAACTAATG'GAGCCTGGCTTTGTTTTTGTTTTATTTCCAACCCACTTGAGCACACAGCACACACAGAGAGAAAAATCAATACTCGTTATGGGATTAAATTTACAAAGCGCAAAGCAAAGCGACAAACAAAATTCAAAAGAAAGAAAAAAAAACACTCAAATAAACTCACAAAGAATTCCTTATCGCCAAGGGGGCCAATGTTCTAAGGTTCTTTCGCCTTGAGAACTTTGAGCTTCCTCTGGCAAAGGAGATTATAATGTACAAATAATGTTGCAATAACCAGTTGAAACCAATGGAATACCGAATCTTGCTAATTAGCAAGGACATCTGTTCACATCTTACCGGGCAGCATTAGATCCTTTTTATAACTCTAATACTGTCAGGTAAAGATGTCGTCCGTGTCCTTAACCTTCAGTACCACCAACAGCAGCAGCAGCACCAAAAAAAAAAAAAAAAAAATGCGTAAAAATCCAAACAAATCATAAAAGTCGAAGGA HsmiPEP155 Homo sapiensGCCGAGCCCGGGCCCAGCGCCGCCTGCAGCCTCGGGAAGGGAGCGGATA SEQ ID NO: 357GCGGAGCCCCGAGCCGCCCGCAGAGCAAGCGCGGGGAACCAAGGAGACGCTCCTGGCACTGCAGATAACTTGTCTGCATTTCAAGAACAACCTACCAGAGACCTTACCTGTCACCTTGGCTCTCCCACCCAATGGAGATGGCTCTAATGGTGGCACAAACCAGGAAGGGGAAATCTGTGGTTTAAATTCTTTATGCCTCATCCTCTGAGTGCTGAAGGCTTGCTGTAGGCTGTATGCTGTTAATGCTAATCGTGATAGGGGTTTTTGCCTCCAACTGACTCCTACATATTAGCATTAACAGTGTATGATGCCTGTTACTAGCATTCACATGGAACAAATTGCTGCCGTGGGAGGATGACAAAGAAGCATGAGTCACCCTGCTGGATAAACTTAGACTTCAGGCTTTATCATTTTTCAATCTGTTAATCATAATCTGGTCACTGGGATGTTCAACCTTAAACTAAGTTTTGAAAGTAAGGTTATTTAAAAGATTTATCAGTAGTATCCTAAATGCAAACATTTTCATTTAAATGTCAAGCCCATGTTTGTTTTTATCATTAACAGAAAATATATTCATGTCATTCTTAATTGCAGGTTTTGGCTTGTTCATTATAATGTTCATAAACACCTTTGATTCAACTGTTAGAAATGTGGGCTAAACACAAATTTCTATAATATTTTTGTAGTTAAAAATTAGAAGGACTACTAACCTCCAGTTATATCATGGATTGTCTGGCAACGTTTTTTAAAAGATTTAGAAACTGGTACTTTCCCCCAGGTAACGATTTTCTGTTCAGGCAACTTCAGTTTAAAATTAATACTTTTATTTGACTCTTAAAGGGAAACTGAAAGGCTATGAAGCTGAATTTTTTTAATGAAATATTTTTAACAGTTAGCAGGGTAAATAACATCTGACAGCTAATGAGATATTTTTTCCATACAAGATAAAAAGATTTAATCAAAAAATTTCATATTTGAAATGAAGTCCCAAATCTAGGTTCAAGTTCAATAGCTTAGCCACATAATACGGTTGTGCGAGCAGAGAATCTACCTTTCCACTTCTAAGCCTGTTTCTTCCTCCATATGGGGATAATACTTTACAAGGTTGTTGTGAGGCTTAGATGAGATAGAGAATTATTCCATAAGATAATCAAGTGCTACATTAATGTTATAGTTAGATTAATCCAAGAACTAGTCACCCTACTTTATTAGAGAAGAGAAAAGCTAATGATTTGATTTGCAGAATATTTAAGGTTTGGATTTCTATGCAGTTTTTCTAAATAACCATCACTTACAAATATGTAACCAAACGTAATTGTTAGTATATTTAATGTAAACTTGTTTTAACAACTCTTCTCAACATTTTGTCCAGGTTATTCACTGTAACCAAATAAATCTCATGAGTCTTTAGTTGATTTAAAATAAAAAAAAAAAAAAAAAAAAAAAAAA AA AtmiPEP157cArabidopsis  CTTTGTCACTTCATACACTCCCTATTGTCTATATATATATATATACTTACASEQ ID NO: 400 thalianaCATATTCAAACATTATAATACTTAATTACACATACATACTTTATGATGTTGCATATCACACATAGGTTTGAGAGTGATGTTGGTTGTTGACAGAAGATAGAGAGCACTAAGGATGACATGCAAGTACATACATATATATCATCACACCGCATGTGGATGATAAAATATGTATAACAAATTCAAAGAAAGAGAGGGAGAGAAAGAGAGAGAACCTGCATCTCTACTCTTTTGTGCTCTCTATACTTCTGTCACCACCTTTATCTCTTCTTCTCTCTAACCT AtmiPEP157d Arabidopsis ATTTACTCTTCACCGCCCTCTCTCTATATATAGTCTCTATCCTCACATATT SEQ ID NO: 401thaliana ATATATCAAACCGCAAGAATGCTGTATGTATAGTGGAGGGTGATAGTGTGGTTGCTGACAGAAGATAGAGAGCACTAAGGATGCTATGCAAAACAGACACAGATATGTGTTTCTAATTGTATTTCATACTTTAACCTCAAAGTTGATATAAAAAAAGAAAGAAAGATAGAAGAGCTAGAAGACTATCTGCATCTCTATTCCTATGTGCTCTCTATGCTTCTGTCATCACCTTTCTTTCTCTATTTCTCTC TAC AtmiPEP160cArabidopsis  AACCAAAACTCTTCAACATTTCTCTCTGACTACTTCATTTCCTCTTCCCAASEQ ID NO: 402 thalianaCAGTTAAAAAAAGTTCTGATTCGATTCAAGCCAAGATCCACGTATAAAGATATGTTCATGCGTAGAGGTTTGGTATACAACAATATATACATATAATAGTTTGTCGTTATGCCTGGCTCCCTGTATGCCACGAGTGGATACCGATTTTGGTTTTAAAATCGGCTGCCGGTGGCGTACAAGGAGTCAAGCATGAC AtmiPEP164b Arabidopsis ATACATTCTCTCTTTCTCTCTCTCTCTCTCTCTCATCCCGGCCCAGTTATGT SEQ ID NO: 403thaliana GGTCGGAGAGAATGATGAAGGTGTGTGATGAGCAAGATGGAGAAGCAGGGCACGTGCATTACTAGCTCATATATACACTCTCACCACAAATGCGTGTATATATGCGGAATTTTGTGATATAGATGTGTGTGTGTGTTGAGTGTGATGATATGGATGAGTTAGTTCTTCATGTGCCCATCTTCACCATC AtmiPEP166c Arabidopsis TCACACATACCTTTCTTTCTCTTCTTCTTCTTACGAAAAGTTTCATCACAT SEQ ID NO: 404thaliana TCACATTATCTTTAACTTTGGTCTCTTTTCTTTTTTGTCTCTTTTCTCTTCTTGATAACGTGGTTCTAGTCTTGATTAATTCATTGTTGTGCGATTTAGTGTTGAGAGGATTGTTGTCTGGCTCGAGGTCATGAAGAAGAGAATCACTCGAATTAATTTGGAAGAACAAATTAAGAAAACCCTAGATGATTCTCGGACCAGGCTTCATTCCCCCTAACCTACTTATCGC AtmiPEP166d Arabidopsis ATTTAGCTTCTTCTTCTTCTTCTTCTTCTGTCTACTTACATAAAGTTATCCTt SEQ ID NO: 405thaliana GCTTTGGTTTAGGGTTGAGAGGAATATTGTCTGGCTCGAGGTCATGAAGAAGATCGGTAGtATTGATTCATTTTAAAGAGTGAAATCCCTAAATGATTCTCGGACCAGGCTTCATTCCCCCCAACC AtmiPEP169a Arabidopsis TAGTATTCATAAGCACCAAAACAAATATGTAGAGATCTCCTCTTCCATTC SEQ ID NO: 406thaliana TCTATTGTTACTTTCGAGAAGAAACATACAAAACACATACATTTTTCTTTTGTTTGTGGTTTTCATATATACAAGTGGGTATAGCTAGTGAAACGCGAATGTGACGAAAGTAGTGTGCAGCCAAGGATGACTTGCCGATTTAAATGATCTTTCTTTATACTCTATTAAGACAATTTAGTTTCAAACTTTTTTTTTTTTTTTTTTTTGAAGGATTCAGGAAGAAATTAGGATATATTATTCCGTATAAAATACAAGATATATAAAACCAAAAAGAAAAAGTAACATGATCGGCAAGTTGTCCTTGGCTACACGTTACTTTGTGTCGC AtmiPEP169h1 Arabidopsis ACTCATCAACAACCTCTTCATAAATACATAAATCATATAAGAGAAAATG SEQ ID NO: 407AtmiPEP169h2 thaliana GTGACATGAAGAATGAGAACTTGTGTGG AtmiPEP169nArabidopsis  AGGCAAAAACATATAGAGAGTAATGAAGTGTATGATGAAGAAGAGAGGSEQ ID NO: 408 thalianaTCTAACATGGCGGAAAGCGTCATGTTTAGTAGCCAAGGATGACTTGCCTGATCTTTTTCGCCTCCACGATTCAATTTCAAATTCATGCATTTTGGATTATTATACCTTTTAAAGTATAATAGGTCAAATATCATGTTGAATCTTGCGGGTTAGGTTTCAGGCAGTCTCTTTGGCTATCTTGACATGCTTTTTCCATCCAT AtmiPEP170Arabidopsis  ATTCACTCCCTTCTTCTTCTTAATCTCCTTACAGTTACAGACATTCTCTCASEQ ID NO: 409 thalianaCTTGCGTTCTTGTTTCTTTTACAAAACAGATACACTATGTTTCCGAGAGAGTCCCTCTGATATTGGCCTGGTTCACTCAGATTCTCTTTTACTAACTCATCTGATTGAGCCGTGTCAATATCTCAGTCCTCTCTCG AtmiPEP396a Arabidopsis TCTCACAACTTCAACTTCCCTCTTTCTCTATATTACGCTTTTGCCCCTCACT SEQ ID NO: 410thaliana CCCTCTTTCCACAATTAGGGTTTCGTCTGCTCTACATGACCCTCTCTGTATTCTTCCACAGCTTTCTTGAACTGCAAAACTTCTTCAGATTTTTTTTTTTTTCTTTTGATATCTCTTACGCATAAAATAGTGATTTTCTTCATATCTCTGCTCGATTGATTTGCGGTTCAATAAAGCTGTGGGAAGATACAGAC AtmiPEP399c Arabidopsis GAATAACCAACCAGCCTTCTCTCAAAGCAAACCAAAAAGAAAAACCAAC SEQ ID NO: 411thaliana ATTGAAAGAGGAAGTTACGATAAGCGGAGCAGTAATAGGGCATCTTTCTATTGGCAGGCGACTTGGCTATTTGTATCTTTTGTGTTCTTGACTATTGGCTATGTCACTTGCCAAAGGAGAGTTGCCCTGTCACTGCTTCCGCTTAAACACAGTCTATAACCGGTTCTGCTAATATCAATCCTTCTTTTGGACATGTCCAAAGCCGAGATTGATTGATAGAGAATTGGTCTCTCTGGCTACAAAACTAGTGCGGTTCTCTCGATTTAAGTTTTAATAGCATTCACTTTGCACATTGCATCTTTCACATCAAATTTCCATTTCATCAACCATCTAAACCTCTTTGTTAGCTTTGATATAAGCAACGATCTAAAGTCTAAAAACCATTAATCCTCTGAAAAAAAAGACAATTTCGATGGTTCTATTATGTTTCTCCAATGCAGAAATTGTATCGTCTGAATTATAGTAGATTTTTTCTAGACTAAAGTGTAAACCAAGACGAATCTGCACTAACAAGACACACCAATAGACTTTACAGAGAAAGGTTACGAGTTTTGAAAATATTAACGGACCATAGTCATCGCG MtmiPEP169d Medicago TGGATAAGTTAATTTATGGTCAAAGAGTCTTTCATGGAGAGGTTGAAAGT SEQ ID NO: 426truncatula GAGATGAAGCCAAGGATGACTTGCCGGTATAATAGTAATTTGCCACAAATCTAGATAGCTATTAGCTATGTTTGGATGGGCGGTGAGATTAACAAAATTACAGCAGCATTGTGATTTTGTTGATGCTTTAAAGTGTAGTTTTTATCAAAATTACAGTGGTTCACTGTAATTATGAGAATCTCACCGTCAATCTAAATATGCATTTAGTTTCATTTCCGGCAGGTCATCCTTCGGCTATATT

TABLE 4 List of the microRNAs (miRNAs) miPEP OrganismSequence of the miRNA SEQ ID AtmiPEP156a1 Arabidopsis thalianaugacagaagagagugagcac SEQ ID NO: 282 AtmiPEP156a2 AtmiPEP156a3AtmiPEP156c1 Arabidopsis thaliana ugacagaagagagugagcac SEQ ID NO: 283AtmiPEP156c2 AtmiPEP156e1 Arabidopsis thaliana ugacagaagagagugagcacSEQ ID NO: 284 AtmiPEP156f1 Arabidopsis thaliana ugacagaagagagugagcacSEQ ID NO: 285 AtmiPEP159a Arabidopsis lyrata uuuggauugaagggagcucuaSEQ ID NO: 286 AtmiPEP159a1 Arabidopsis thaliana uuuggauugaagggagcucuaSEQ ID NO: 287 CrmiPEP159a Capsella rubella uuuggauugaagggagcucuaSEQ ID NO: 288 AtmiPEP159b1 Arabidopsis thaliana uuuggauugaagggagcucuuSEQ ID NO: 289 AtmiPEP159b2 AtmiPEP160a1 Arabidopsis thalianaugccuggcucccuguaugcca SEQ ID NO: 290 AtmiPEP160b1 Arabidopsis thalianaugccuggcucccuguaugcca SEQ ID NO: 291 AtmiPEP160b2 AtmiPEP161Arabidopsis thaliana ucaaugcauugaaagugacua SEQ ID NO: 292 AtmiPEP162a1Arabidopsis thaliana ucgauaaaccucugcauccag SEQ ID NO: 293 AtmiPEP162b1Arabidopsis thaliana ucgauaaaccucugcauccag SEQ ID NO: 294 AtmiPEP163-1Arabidopsis thaliana uugaagaggacuuggaacuucgau SEQ ID NO: 295AtmiPEP163-2 AtmiPEP164a1 Arabidopsis lyrata uggagaagcagggcacgugcaSEQ ID NO: 296 AtmiPEP164a2 AtmiPEP164a3 AtmiPEP164a1Arabidopsis thaliana uggagaagcagggcacgugca SEQ ID NO: 297 AtmiPEP164a2AtmiPEP164a3 BrmiPEP164a1 Brassica rapa uggagaagcagggcacgugcaSEQ ID NO: 298 BrmiPEP164a2  BrmiPEP164a3 CpmiPEP164a1 Carica papayauggagaagcagggcacgugca SEQ ID NO: 299 CpmiPEP164a2 CrniPEP164a1Capsella rubella uggagaagcagggcacgugca SEQ ID NO: 300 CrmiPEP164a2 CrmiPEP164a3 GrmiPEP164a1 Gossypium raimondii uggagaagcagggcacgugcaSEQ ID NO: 301 GrmiPEP164a2  GrmiPEP164a3 MtmiPEP164a1Medicago truncatula uggagaagcagggcacgugca SEQ ID NO: 302 MtmiPEP164a2OsmiPEP164a1 Oryza sativa uggagaagcaggguacgugca SEQ ID NO: 303OsmiPEPl64a2 AlmiPEP165a Arabidopsis lyrata ucggaccaggcuucaucccccSEQ ID NO: 304 AtmiPEP165a Arabidopsis thaliana ucggaccaggcuucaucccccSEQ ID NO: 305 BcmiPEP165a Brassica carinata ucggaccaggcuucaucccccSEQ ID NO: 306 BjmiPEP165a Brassica juncea ucggaccaggcuucaucccccSEQ ID NO: 307 BnmiPEP165a Brassica napus ucggaccaggcuucaucccccSEQ ID NO: 308 BomiPEP165a Brassica oleracea ucggaccaggcuucaucccccSEQ ID NO: 309 BrmiPEP165a Brassica rapa ucggaccaggcuucaucccccSEQ ID NO: 310 AtmiPEP166a Arabidopsis thaliana ucggaccaggcuucauuccccSEQ ID NO: 311 AtmiPEP166b Arabidopsis thaliana ucggaccaggcuucauuccccSEQ ID NO: 312 AtmiPEP167a Arabidopsis thaliana ugaagcugccagcaugaucuaSEQ ID NO: 313 AtmiPEP167b1 Arabidopsis thaliana ugaagcugccagcaugaucuaSEQ ID NO: 314 AtmiPEP167b2 AtmiPEP169c1 Arabidopsis thalianacagccaaggaugacuugccgg SEQ ID NO: 315 AtmiPEP169c2 AtmiPEP16911Arabidopsis thaliana uagccaaggaugacuugccug SEQ ID NO: 316 AtmiPEP171a1Arahidopsis thaliana ugauugagccgcgccaauauc SEQ ID NO: 317 AtmiPEP171bArabidopsis thaliana uugagccgugccaauaucacg SEQ ID NO: 318 MtmiPEP171b1Medicago truncatula ugauugagccgcgucaauauc SEQ ID NO: 319 MtmiPEP171b2ZmmiPEP171b Zea mays ggauugagccgcgucaauauc SEQ ID NO: 320 AtmiPEP171c1Arabidopsis thaliana uugagccgugccaauaucacg SEQ ID NO: 321 MtmiPEP171cMedicago truncatula agauugagccgcgccaauauc SEQ ID NO: 322 MtmiPEP171hMedicago truncatula cgagccgaaucaauaucacuc SEQ ID NO: 323 AtmiPEP172a1Arabidopsis thaliana agaaucuugaugaugcugcau SEQ ID NO: 324 AtmiPEP172a3AtmiPKP172b1 Arabidopsis thaliana gcagcaccauuaagauucac SEQ ID NO: 325AtmiPEP172c1 Arabidopsis thaliana agaaucuugaugaugcugcag SEQ ID NO: 326AtmiPEP172e1 Arabidopsis thaliana ggaaucuugaugaugcugcau SEQ ID NO: 327AtmiPEP172e2 AtmiPEP172e3 AcmiPEP319a1 Arabidopsis cebennensisuuggacugaagggagcucccu SEQ ID NO: 328 AcmiPEP319a2 AhmiPEP319aArabidopsis halleri uuggacugaagggagcucccu SEQ ID NO: 329 AlmiPKP319aArabidopsis lyrata uuggacugaagggagcucccu SEQ ID NO: 330 AtmiPEP319a1Arabidopsis thaliana uuggacugaagggagcucccu SEQ ID NO: 331 AtmiPEP319a2BrmiPEP319a Brassica rapa uuggacugaagggagcucccu SEQ ID NO: 332CpmiPEP319a Carica papaya uuggacugaagggagcuccuu SEQ ID NO: 333CrmiPEP319a Capsella rubella uuggacugaagggagcucc SEQ ID NO: 334EgmiPEP319a Eucalyptus grandis uuggacugaagggagcucccu SEQ ID NO: 335GrmiPEP319a Gossypium raimondii uuggacugaagggagcucccu SEQ ID NO: 336MtmiPEP3l9a Medicago truncatula uuggacugaagggagucucccu SEQ ID NO: 337OsmiPEP319a Oryza saliva uuggacugaagggugcucccu SEQ ID NO: 338PpmiPEP319a Physccomitrella patens cuuggacugaagggagcucc SEQ ID NO: 339ThmiPEP319a1 Thellungiella halophila uggacucaaggaagcucucu SEQ ID NO: 340ThmiPEP319a2 AtmiPEP319b1 Arabidopsis thaliana uuggacugaagggagcucccuSEQ ID NO: 341 AtmiPEP394a1 Arabidopsis thaliana uuggcauucuguccaccuccSEQ ID NO: 342 AtmiPEP395c1 Arabidopsis thaliana cugaaguguuuggggggacucSEQ ID NO: 343 AtmiPEP395c1 Arabidopsis thaliana cugaaguguuugggggaacucSEQ ID NO: 344 AtmiPEP397b1 Arabidopsis thaliana ucauugagugcaucguugaugSEQ ID NO: 345 AtmiPEP398c1 Arabidopsis thaliana uguguucucaggucaccccugSEQ ID NO: 346 AtmiPEP399b Arabidopsis thaliana ugccaaaggagaguugcccugSEQ ID NO: 347 AtmiPEP399d1 Arabidopsis thaliana ugccaaaggagauuugccccgSEQ ID NO: 348 AtmiPEP403 Arabidopsis thaliana uuagauucacgcacaaacucgSEQ ID NO: 349 AtmiPEP447a1 Arabidopsis thaliana uuggggacgagauguuuuguugSEQ ID NO: 350 AtmiPEP447a2 Arabidopsis thaliana uuggggacgagauguuuuguugAtmiPEP447b1 Arabidopsis thaliana uuggggacgagauguuuuguug SEQ ID NO: 351AtmiPEP447b2 Arabidopsis thaliana uuggggacgagauguuuuguug AtmiPEP447cArabidopsis thaliana ccccuuacaauguc gaguaaa SEQ ID NO: 352 DmmiPEP1aDrosophila melanogaster uggaauguaaagaaguauggag SEQ ID NO: 353 DmmiPEP1bDmmiPEP8 Drosophila melanogaster uaauacugucagguaaagauguc SEQ ID NO: 354HsmiPEP155 Homo sapiens uuaaugcuaaucgugauaggggu SEQ ID NO: 358AtmiPEP157c Arabidopsis thaliana uugacagaagauagagagcac SEQ ID NO: 412AtmiPEP157d Arabidopsis thaliana ugacagaagauagagagcac SEQ ID NO: 413AtmiPEP160c Arabidopsis thaliana ugccuggcucccuguaugcca SEQ ID NO: 414AtmiPEP164b Arabidopsis thaliana uggagaagcagggcacgugca SEQ ID NO: 415AtmiPEP166c Arabidopsis thaliana ucggaccaggcuucauucccc SEQ ID NO: 416AtmiPEP166d Arabidopsis thaliana ucggaccaggcuucauucccc SEQ ID NO: 417AtmiPEP169a Arabidopsis thaliana cagccaaggaugacuugccga SEQ ID NO: 418AtmiPEP169h Arabidopsis thaliana uagccaaggaugacuugccug SEQ ID NO: 419AtmiPEP169n Arabidopsis thaliana uagccaaggaugacuugccug SEQ ID NO: 420AtmiPEP170 Arabidopsis thaliana ugauugagccgugucaauauc SEQ ID NO: 421AtmiPEP396a Arabidopsis thaliana uuccacagcuuucuugaacug SEQ ID NO: 422AtmiPEP399c Arabidopsis thaliana ugccaaaggagaguugcccug SEQ ID NO: 423AtmiPEP169d Medicago truncatula aagccaaggaugacuugccgg SEQ ID NO: 427

TABLE 5 List of the control Pre-miRNAs Pre-miRNA OrganismSequence of the Pre-miRNA SEQ ID Pre-miR169 MedicagoTTAGGGTTTTCAGCTCATGGTAATAAAAATGTCATCTAATGTCTTGCATGT SEQ ID NO: 359truncatula GGGAATGAGGTCATATATGCAGCCAAGGATGACTTGCCGGCGAGCCTCTTTCGATACTTTTATGACATAATTAATCATGTGGATAGCCAAGGTACTAAACTCACTTTGCACTAAAACAAATATTTTTGCTTTAGTGCAAACTTAGTTTAGGCGCTTCGCAACGGCTAGTCAAATGTCCTAGTTCCAATGTGATTGGTTGTCCGGCAAGTCGTCTCTGGCTACGTAAAGGCCTCCTTTTTTCATGCTAGATTTTTGATGATTTGATATAGCCACACATATTTTGGAA Pre-miR169a Medicago AAGAGGCAGAGAGAGTAATGCAGCCAAGGATGACTTGCCGACAACATTG SEQ ID NO: 360truncatula GCGAATGTTCATGTGATTTCTGCCTCATTGTGCCGGCAAGTTGTCCTTGGCTATGTTAGTCTCTCATCTTCT Pre-miR171a  Medicago TGAATTCCCCTCCGCTTTTTGATGTTGGCTTGTCTCAATCAAATCAAAGTTC SEQ ID NO: 361MI0001753 truncatula TTGAAATTTGAGTTCTTTAGTCTGATTGAGTCGTGCCAATATCATATTAAGCGATAAAAGTC Pre-miR171h Medicago CCACAAAACTATAACTAGCTAGAAGCTTTAATCGCCTTATTTATTATAATA SEQ ID NO: 362truncatula ATAATAATAAATATGGCTTCAGCTGCAAAAGTATACATGGCGTGATATTGATCCGGCTCATCTATATCTTCAAGTTCAATCATCCATATTCATATCAATTTCAGACGAGCCGAATCAATATCACTCTTGTTTGCTTCATTGCATATTAATTATATACTTCATTTATAAGTTATAGTTTGCCATATATATATTAGATTGATTCTGCAGAAGTAGACAGGAGTGGTGTTGTTTCTGCTCATCTTATTAAATAATGAATGAATGAATGACATTTGCTTACTTATAAGACGAGCCGAATCAATATCACTCC AGTACACCTPre-miR393a Medicago  AACTGCAACTTGAGGAGGCATCCAAAGGGATCGCATTGATCCTATAATATSEQ ID NO: 363 truncatulaTTCAACTTTAGTCACTTTAATTTTCTCTCATATAATACTTAATTGGGATCATGCCATCCCTTTGGATTTCTCCTTTAGTAGCTAC Pre-miR393b Medicago AGGCATCCAAAGGGATCGCATTGATCCCAAATCTAATTAAGTCCCTAGCTA SEQ ID NO: 364truncatula CTTAATTAACAACTTAATTTCCTTAATATCTCATAATATTTGGGATCATGCTATCCCTTTGGATTCAT Pre-miR396a Medicago TGCTTTTCCACAGCTTTCTTGAACTTCTTTCGTATCTTAAATCTGTTTTCAA SEQ ID NO: 365truncatula GATTAAAAGTCCTAGAAGCTCAAGAAAGCTGTGGGAGAATA Pre-miR396bMedicago  TATTCTTCCACAGCTTTCTTGAACTGCATCCAAATTGAGTTCCTTTGCATTGSEQ ID NO: 366 truncatula CCATGGCCATTGTTTTGCGGTTCAATAAAGCTGTGGGAAGATA

TABLE 6 List of the control miRNAs miRNA Organism Sequence of the miRNASEQ ID miR169 Medicago truncatula CAGCCAAGGAUGACUUGCCGG SEQ ID NO: 367miR169a Medicago truncatula CAGCCAAGGAUGACUUGCCGA SEQ ID NO: 368 miR171aMedicago truncatula UGAUUGAGUCGUGCCAAUAUC SEQ ID NO: 369 miR171hMedicago truncatula GAGCCGAAUCAAUAUCACUC SEQ ID NO: 370 miR393aMedicago truncatula UCCAAAGGGAUCGCAUUGAUC SEQ ID NO: 371 miR393bMedicago truncatula UCCAAAGGGAUCGCAUUGAUC SEQ ID NO: 372 miR396aMedicago truncatula UUCCACAGCUUUCUUGAACUU SEQ ID NO: 373 miR396bMedicago truncatula UUCCACAGCUUUCUUGAACUG SEQ ID NO: 374

TABLE 7 Polymorphism of the DNA sequence of the different regions ofpri-miR171b # # % # Size SNPs mutations SNP haplotypes pri-mir171b 112791 100 8.07 161 5′ pri-miR171b 129 4 4 3.1 5 miPEP171b 62 2 2 3.22 3Pre-miR171b 118 1 1 0.85 2 miR171b + miR171b* 42 0 0 0 1 3′ pri-miR171b259 39 42 15.06 89

The following examples will illustrate the invention better, but withoutlimiting its scope.

EXAMPLES

A: Analysis of the miPEPS in Plants

Example 1 Characterization in the Model Plant Medicago truncatula

a) Identification and characterization of MtmiPEP171b1 (miPEP171b1identified in Medicago truncatula)

The microRNA MtmiR171b is expressed in the meristematic region of theroots. The overexpression of this microRNA in particular leads to areduction in the expression of the genes HAM1 (Accession No.MtGI9-TC114268) and HAM2 (Accession No. MtGI9-TC120850) (FIG. 1A), aswell as to a reduction in the number of lateral roots (FIG. 1B). Theoverexpression of the pri-miR171b also induces a decrease in the numberof lateral roots (FIG. 37).

The sequence of the primary transcript of MtmiR171b was determined usingthe RACE-PCR technique. Analysis of the sequence of the primarytranscript made it possible to identify the presence of severalcompletely unexpected small open reading frames (sORFs). These sORFswere called miORFs for “microRNA ORFS”. These miORFs potentially encodeshort peptides, from about 4 to 100 amino acids. No significant homologyrelating to these miORFS was found in the databases.

The overexpression of the first miORF, called MtmiORF171b, leads to anincrease in the accumulation of MtmiR171b and a reduction in theexpression of the HAM1 and HAM2 genes (see FIG. 2A), as well as to areduction in the number of lateral roots (FIG. 2B), as was alreadyobserved in the overexpression of MtmiR171b.

In order to determine whether MtmiORF171b leads to the real productionof a peptide and whether the regulatory function observed above isindeed carried by said peptide, a synthetic peptide, the sequence ofwhich is identical to that potentially encoded by MtmiORF171b, wasapplied on the roots of Medicago truncatula. Application of this peptideleads to the phenotype already observed above in the overexpression ofMtmiORF171b, i.e. it leads to an increase in the accumulation ofMtmiR171b and a reduction in the expression of the HAM1 and HAM2 genes(see FIG. 3A), as well as a reduction in the number of lateral roots(FIG. 3B).

The results of these experiments demonstrate that MtmiORF171b encodes apeptide capable of modulating the accumulation of MtmiR171b, and theexpression of the target genes of MtmiR171b: HAM1 and HAM2. Said peptidehas been called MtmiPEP171b1 (“miPEP” corresponding to microRNA encodedPEPtide).

Moreover, MtPEP171b1 leads to an increase in the accumulation ofMtmiR171b (FIG. 4A) and of pre-MtmiR171b (FIG. 4B).

b) Specificity of miPEP171b1

The expression of different microRNA precursors of Medicago truncatula(MtmiR171b SEQ ID NO: 319, MtmiR169 SEQ ID NO: 367, MtmiR169a SEQ ID NO:368, MtmiR171a SEQ ID NO: 369, MtmiR171h SEQ ID NO: 370, MtmiR393a SEQID NO: 371, MtmiR393b SEQ ID NO: 372, MtmiR396a SEQ ID NO: 373 andMtmiR396b SEQ ID NO: 374) was determined and compared between controlplants and plants in which MtmiORF171b encoding MtmiPEP171b1 wasoverexpressed (FIG. 5A), or between control plants and plants grown onculture medium containing MtmiPEP171b1 (FIG. 5B).

The results obtained indicate that MtmiPEP171b1 only leads to anincrease in the accumulation of MtmiR171b and not of the other miRNAs,which indicates that a miPEP only has an effect on the microRNA fromwhich it is derived.

c) Localization of miPEP171b1

Moreover, the immunolocalization of miPEP171b1 in the roots of M.truncatula reveals the presence of miPEP171b1 in the lateral rootinitiation sites, showing a co-localization between the microRNA and thecorresponding miPEP (FIGS. 28 and 34).

Example 1B MtmiPEP169d

Regarding MtmiPEP169d, it has been demonstrated in vivo in M. truncatulathat the overexpression of this miPEP induces an accumulation ofMtmiR169d (FIG. 32).

Example 1C MtmiPEP171e

Regarding MtmiPEP171e, it has been demonstrated in vivo in M. truncatulathat the overexpression of this miPEP induces an accumulation ofMtmiR171e (FIG. 33).

Example 2 Characterization in the Model Plant of Tobacco

a) Conservation of the Mechanism in Tobacco

In order to determine whether the mechanism of regulation of themicroRNAs is conserved in other plant species, the regulation ofMtmiR171b by MtmiPEP171b1 was tested in a different cellular model. Forthis, MtmiR171b and MtmiPEP171b1 were introduced into tobacco leaves.

The accumulation of MtmiR171b was measured in tobacco leaves transformedin order to express MtmiR171b of Medicago truncatula starting fromwild-type pri-miRNA capable of producing MtmiPEP171b1, or starting froma mutated version of pri-miRNA incapable of producing MtmiPEP171b1 (inwhich the start codon ATG of MtmiORF171b has been replaced with ATT)(FIG. 6 and FIG. 20). Absence of translation of MtmiPEP171b1 leads to amarked decrease in the accumulation of MtmiR171b.

The accumulation of pre-MtmiR171b was measured in tobacco leavestransformed in order to express MtmiR171b of Medicago truncatula alone(control), or additionally expressing the wild-type MtmiORF171b ofMedicago truncatula (35SmiPEP171b1 ATG), or a mutated version ofMtmiORF171b in which the start codon ATG has been replaced with ATT(35SmiPEP171b1 ATT) (FIG. 7 and FIG. 21). The expression of MtmiORF171bleads to an increase in the accumulation of pre-miR171b, and thisaccumulation of pre-miR171b depends on the translation of MtmiORF171b tomicropeptide.

Moreover, in the tobacco leaves transformed in order to expressMtmiR171b of Medicago truncatula, untreated or treated by spraying withMtmiPEP171b1 (0.1 μM) for the first time 12 h before sampling and then asecond time 30 minutes before sampling, it was observed thatMtmiPEP171b1 may be used directly in peptide form by foliar sprayings(FIG. 8).

Moreover, it was observed in tobacco (as in Medicago truncatula) thatthe MtmiPEP171b1 leads to an increase in the accumulation of MtmiR171b(FIG. 9A) and of pre-MtmiR171b (FIG. 9B), but reduces the accumulationof pri-MtmiR171b (FIG. 9C).

Taken together, these results indicate that the mechanism of regulationof the microRNAs and of their target genes under the control of miPEPsis conserved between the species.

b) Intracellular Localization of MtmiPEP171b1

Tobacco leaves were transformed in order to overexpress MtmiPEP171b1 ofMedicago truncatula fused with a fluorescent protein (GFP) (FIG. 10).The results obtained indicate that the miPEP is localized in smallnuclear bodies.

c) Identification of miPEPS from Databases

Genomic databases of plants were searched for the presence of openreading frames within primary transcripts of 70 miRNAs, and 101 miORFscapable of encoding a miPEP were identified.

At present, AtmiPEP165a and AtmiPEP319a2, identified in Arabidopsisthaliana, have already been characterized. The experiments conducted inthe model plant of tobacco made it possible to demonstrate that theoverexpression of AtmiORF165a or of AtmiORF319a leads to an increase inthe accumulation of AtmiR165a or of AtmiR319a respectively (FIG. 11).

miR165a regulates transcription factors such as Revoluta, Phavoluta andPhabulosa.

miR319 regulates genes of the TCP family.

Example 3 Characterization in the Arabidopsis thaliana Model PlantExample 3A AtmiPEP165a

Regarding AtmiPEP165a, it has been demonstrated in vivo in Arabidopsisthaliana that treatment with AtmiPEP165a leads to a phenotype withgreatly accelerated root growth (FIG. 12).

Moreover, treatment of plants with higher and higher concentrations ofmiPEP165a shows a dose-dependent effect on the accumulation of miR165aand the negative regulation of its target genes (PHAVOLUTA: PHV,PHABOLUSA: PHB and REVOLUTA: REV) as a function of the amount ofmiPEP165A (see FIG. 22).

Example 3B AtmiPEP164a

Regarding AtmiPEP164a, this was synthesized and was used forinvestigating an increase in the accumulation of miR164a in roots of A.thaliana treated with the synthetic peptide.

Northern blot analyses indicate that treatment of the plant with thepeptide miPEP164a leads to an increase in the accumulation of miR164a(FIG. 23).

The same type of results has been obtained by qRT-PCR (FIG. 30).

It has also been demonstrated in vivo in Arabidopsis thaliana thattreatment of the plant with AtmiPEP164a increases plant growthsignificantly (FIG. 24).

Example 3C AtmiPEP165a

Regarding AtmiPEP165a, this was synthesized and was used forinvestigating an increase in the accumulation of miR165a in roots of A.thaliana treated with the synthetic peptide.

Northern blot analyses indicate that treatment of the plant with thepeptide miPEP165a leads to an increase in the accumulation of miR165a(FIG. 25).

Moreover, to determinate the mode of action of miPEPs regarding theaccumulation of their own miRs, the expression of pri-miR165a has beenanalysed in wildtype plants and in plants that have been transformed tooverexpress miPEP165a (FIG. 35).

The overexpression of pri-miR165a is enhanced in the plants thatoverexpress miPEP165a (FIG. 36A).

In the presence of cordycepin, an inhibitor of RNA synthesis, the amountof pri-miR165a is identical in wildtype plants and in transformed plants(FIG. 36B), it indicates that miPEP165a does not operate as a RNAstabilizer, but rather as a transcription activator of pri-miR165a.

Moreover, the expression of pri-miR165a has been analysed in wildtypeplants (Col-0 plants) and in mutated plants having a low allele of thesecond large subunit of RNA polymerase II (nrpb2-3 plants). The resultsobtained indicate that mutated plants do not show an increase in theaccumulation of pri-miR165 in response to miPEP165a, unlike wildtypeplants. It appears that the miPEP operates as a transcription regulator.

Example 3D AtmiPEP319a1

Regarding AtmiPEP319a1, this was also synthesized and was used forinvestigating an increase in the accumulation of miR319a in roots of A.thaliana treated with the synthetic peptide.

Analyses by qRT-PCR show that the overexpression of AtmiPEP319a1 leadsto an increase in the accumulation of miR319a (FIGS. 26 and 31).

It was also demonstrated in vivo in Arabidopsis thaliana that treatmentof the plant with AtmiPEP319a1 increases plant growth significantly(FIG. 27).

Example 3E AtmiPEP160b1

Regarding AtmiPEP160b1, it has been demonstrated in vivo in A. thalianathat the overexpression of this miPEP induces an accumulation ofAtmiR160b (FIG. 29).

Material and Methods

Biological Material

The surface of seeds of M truncatula was sterilized and they were leftto germinate on agar plates for 5 days at 4° C. in the dark. The youngshoots were then grown on 12-cm square plates filled with Fahraeusmedium without nitrogen and containing 7.5 μM phosphate (Lauressergueset al., Plant J., 72(3): 512-22, 2012). The lateral roots were countedevery day. In pots, the plants were watered every other day withmodified Long Ashton medium with low phosphorus content (Balzergue etal., Journal of Experimental Botany, (62)1049-1060, 2011).

The peptides were synthesized by Eurogentec or Smartox-Biotech.MtmiPEP171b1 was resuspended in a solution of 40% water/50%acetonitrile/10% acetic acid (v/v/v), and the other peptides wereresuspended in water.

The leaves were watered by spraying with the peptides using peptidesolutions at different concentrations (0.01, 0.1, 1 μM), firstly 12 hbefore sampling and then a second time 30 min before sampling.

Reverse Transcription of the microRNAs

The RNA was extracted using the reagent Tri-Reagent (MRC) according tothe manufacturer's instructions, except for precipitation of the RNA,which was carried out with 3 volumes of ethanol. The eversetranscription of the RNA was carried out using the specific stem-loopprimer RTprimer171b in combination with hexamers for performing thereverse transcription of RNA of high molecular weight.

In brief, 1 μg of RNA was added to the stem-loop primer MIR171b (0.2μM), the hexamer (500 ng), the buffer RT (1×), the enzyme SuperScriptReverse transcriptase (SSIII) (one unit), the dNTPs (0.2 mM each), DTT(0.8 mM) in a final reaction mixture of 25 μl. In order to carry out thereverse transcription, a reaction of pulsed reverse transcription wasperformed (40 repetitions of the following cycle: 16° C. for 2 minutes,42° C. for one minute and 50° C. for one second, followed by a finalinactivation of the reverse transcription at 85° C. for 5 minutes).

Analyses by Quantitative RT-PCR (qRT-PCR)

The total RNA was extracted from roots of M. truncatula or from tobaccoleaves using the extraction kit RNeasy Plant Mini Kit (Qiagen). Thereverse transcription was performed using the reverse transcriptaseSuperScript II (Invitrogen) starting from 500 ng of total RNA. Threerepetitions (n=3) were carried out, each with two technical repetitions.Each experiment was repeated from two to three times. The amplificationsby qPCR were carried out using a LightCycler 480 System thermocycler(Roche Diagnostics) by the method described in Lauressergues et al.(Plant J., 72(3): 512-22, 2012).

Statistical Analyses

The mean values of the relative expression of the genes or of theproduction of lateral roots were analysed using the Student test or theKruskal-Wallis test. The error bars represent the SEM (Standard Error ofthe Mean). The asterisks indicate a significant difference (p<0.05).

Plasmid Constructs

The DNA fragments of interest were amplified with Pfu polymerase(Promega). The DNA fragments were cloned using the XhoI and NotI enzymesinto a pPEX-DsRED plasmid for an overexpression under the control of theconstitutive strong promoter 35S, and using the KpnI-NcoI enzymes into apPEX GUS plasmid for the reporter genes, by the method described inCombier et al. (Genes & Dev, 22: 1549-1559, 2008).

For the miPEPs 165a and 319a, the corresponding miORFs were cloned intopBIN19 by the method described in Combier et al. (Genes & Dev, 22:1549-1559, 2008).

Transformation of the Plants

The composite plants having roots transformed with Agrobacteriumrhizogenes were obtained by the method described in Boisson-Dernier etal. (Mol Plant-Microbe Interact, 18: 1269-1276, 2005). The transformedroots were verified and selected by observations of DsRED with abinocular fluorescence magnifier. The control roots correspond to rootstransformed with A. rhizogenes not containing the pPEX-DsRED vector.Transformation of the tobacco leaves was carried out by the methoddescribed in Combier et al. (Genes & Dev, 22: 1549-1559, 2008).

Northern Blot

Northern blot analysis was carried out according to the protocoldescribed in Lauressergues et al. Plant J., 72(3): 512-22, 2012.

The biological samples were homogenized in a buffer containing 0.1 M ofNaCl, 2% of SDS, 50 mM of Tris-HCl (pH 9), 10 mM of EDTA (pH 8) and 20mM of mercaptoethanol, and the RNA was extracted twice with aphenol/chloroform mixture and was precipitated with ethanol.

The RNA was loaded on PAGE 15% gel and transferred to a nylon membrane(HybondNX, Amersham). RNA was hybridized with a radioactiveoligonucleotide probe labelled at its end, in order to detect the RNA U6or for miR164a.

The hybridizations were carried out at 55° C. The hybridization signalswere quantified using a phosphorimager (Fuji) and normalized with thesignal of the specific probe of RNA U6.

RNA Stability

For each condition, two week old plants that have been cultivatedvertically on solid MS medium have been transferred in 6-well platescontaining 1 ml of liquid MS medium. After a 16 hour incubation with 1mM of miPEP, the plants have been treated with 100 μg/ml of cordycepinor with water (as control), and harvested a different times to extractand quantify RNA. Each indivual experiment has been made 3 times.

Histochemical Labelling

Labelling with GUS was carried out by the method described in Combier etal., (Genes & Dev, 22: 1549-1559, 2008). The samples were observed witha microscope (axiozoom).

Immunolocalization

Roots or plantlets of tissues of Medicago were fixed for 2 hours in 4%formol (v/v) with 50 mM of phosphate buffer (pH 7.2), and then embeddedin agarose LMP 5% in water (with a low melting point). Thin sections(100 μm) were obtained and were placed in Pbi (phosphate buffer forimmunology) on Teflon-coated slides, blocked in Pbi, 2% Tween and 1% ofbovine serum albumin for 2 hours (PbiT-BSA), then labelled overnight (12h) at 4° C. with the primary antibody diluted in BSA-PbiT. The sectionswere washed with PBiT and incubated at ambient temperature for 2 h witha secondary antibody diluted in PbiT-BSA. The slides were then washed inPbi for 30 min and mounted in Citifluor (mounting medium). The primaryantibodies and the dilutions were as follows: 1716a (1:500, v/v). Thesecondary antibody was a goat anti-rabbit IgG antibody coupled to theAlexa Fluor 633 fluorescent probe (Molecular Probes), and was used at adilution of 1:1000 (v/v).

Western Blotting

A total extract of proteins has been prepared using the method describedin Combier et al., (Genes & Dev, 22: 1549-1559, 2008) and separated bySDS-PAGE. The transfer has been made using a phosphate buffer overnightat 4° C., at 15V, then the membrane has been incubated for 45 min atroom temperature in a solution of glutaraldehyde 0.2% (v/v). Primaryantibodies have been used at a 1:1000 (v/v) dilution and HRP-conjugatedgoat anti-rabbit Ig antibodies have used as secondary antibodies at a1:40 000 (v/v) dilution.

TABLE 8 List of primers AtmiR160bq5 (SEQ ID NO: 429)TCCCCAAATTCTTGACCAAA AtmiR160bq3 (SEQ ID NO: 430) TTGAGGGGAAAACATGAACCAtmiR164aq5 (SEQ ID NO: 431) ACGAAATCCGTCTCATTTGCAtmiR164aq3 (SEQ ID NO: 432) GGGTGAAGAGCTCATGTTGGAtmiR165q5 (SEQ ID NO: 433) TGAGGGGAATGTTGTCTGGAtmiR165q3 (SEQ ID NO: 434) GAAGCCTGGTCCGAGGATAAtmiR319q5 (SEQ ID NO: 435) CGAGTCGCCAAAATTCAAACAtmiR319q3 (SEQ ID NO: 436) GCTCCCTTCAGTCCAATCAAAtPHVq5 (SEQ ID NO: 437) GCAACTGCAGTGGAATAGCA AtPHVq3 (SEQ ID NO: 438)GCGACCTTCATGGGTTCTAA AtPHBq5 (SEQ ID NO: 439) CTCAGCATCAGCAACGTGATAtPHBq3 (SEQ ID NO: 440) AACTCTGCTAGGGCCTCCTC AtREVq5 (SEQ ID NO: 441)TCACAACTCCTCAGCATTCG AtREVq3 (SEQ ID NO: 442) ACCCAATCAACAGCAGTTCCAtactine2F (SEQ ID NO: 443) GGTAACATTGTGCTCAGTGGAtactine2R (SEQ ID NO: 444) CTCGGCCTTGGAGATCCACAmiORF319acla5 (SEQ ID NO: 445) tcatcgATGAATATACATACATACCATCATmiORF319abam3 (SEQ ID NO: 446) tccggatccTCTAAAAGCTAAAGTGATTCTAAAAtmiR319cla5 (SEQ ID NO: 447) tcatcgAGAGAGAGCTTCCTTGAGTCAtmiR319bam3 (SEQ ID NO: 448) tccggatccAGAGGGAGCTCCCTTCAGTAtmiR165cla5 (SEQ ID NO: 449) tcatcgGTTGAGGGGAATGTTGTCTGAtmiR165bam3 (SEQ ID NO: 450) tccggatccGTTGGGGGGGATGAAGCCAtmiR165muorf5cla (SEQ ID NO: 451) tcatcgATGAGGGTTAAGCTATTTCAGTAtmiR165muorf3bam (SEQ ID NO: 452) tccggatccTAATATCCTCGATCCAGACAACmiR171bq5 (SEQ ID NO: 453) CGCCTCAATTTGAATACATGGmiR171bq3 (SEQ ID NO: 454) ACGCGGCTCAATCAAACTACpri-miR171bq5 (SEQ ID NO: 455) GCACTAGCTGGTTTCATTATTCCpri-miR171bq3 (SEQ ID NO: 456) TTGCAAAATTTGGAGAGCCTAmiR171eq5 (SEQ ID NO: 457) AAGCGATGTTGGTGAGGTTCmiR171eq3 (SEQ ID NO: 458) CGGCTCAATCTGAGATCGTATmiR169dq5 (SEQ ID NO: 459) GGGCGGTGAGATTAACAAAAmiR169dq3 (SEQ ID NO: 460) CCTGCCGGAAATGAAACTAAmiORF171e2Cla5 (SEQ ID NO: 461) tcatcgATGATGGTGTTTGGGAAGCCmiORF171e2bam3(SEQ ID NO: 462) tccggatccTACATGTAAATCCGTCTTCGGmiORF171b5′AGG (SEQ ID NO: 463) AGGCTTCTTCATAGGCTCTCCmiR171e5′Xho (SEQ ID NO: 464) TCAGTCCGCTCGAGGAATAAGTGAATAT TATCGATATTTmiR171e3'Not (SEQ ID NO: 465) AAGGAAAAAAGCGGCCGCAAAGTGATA TTGGCGCGGCT171b scramble F bis (SEQ ID NO: 466) TAGCATGTTATTACACCGCTTAAGTAAGTTCTGTAAGATAGAGCGCGATATTGTTT ACATTAGCTAG171b scramble R bis (SEQ ID NO: 467) CGTACTAGCTAATGTAAACAATATCGCGCTCTATCTTACAGAACTTACTTAAGCGG TGTAATAACAT PromiORF171b1 ATG BsaIR aaaaGGTCTCgCCATAAGCATAGTGGAAT (SEQ ID NO: 468) AATGAAACCPromiORF171b2 ATG BsaIR  aaaaGGTCTCgCCATTTCCATACCTGAATT (SEQ ID NO: 469)TCTCCTT PromiORF171b6 ATG BsaI R  aaaaGGTCTCgCCATCTTCATCCTCAATAT(SEQ ID NO: 470) CCTAATTC 171b fusion GFP sans ATG BsaIaGGTCTCgACACAGATATATATACTATGT R (SEQ ID NO: 471) CTCTTTCAATTTTGCPro171b fusionTranscript GUSpPEX aaaGGTCTCgCCATGACCATTTTCCCACGBsaI R (SEQ ID NO: 472) AATATG Pro171b fusionATG1aaGGTCTCgACACAAGCATAGTGGAATA GUSpPEXsansATG BsaI R  ATGAAACC(SEQ ID NO: 473) Pro171bfusionTraducGUSpPEXaaGGTCTCgACACAGATATATATACTATG BsaI R (SEQ ID NO: 474) TCTCTTTCA171b NcoI F (SEQ ID NO: 475) Gaattcctgcccatggttttcg171b NcoI R (SEQ ID NO: 476) Cgaaaaccatgggcaggaattc171b AflII F (SEQ ID NO: 477) cagtcccgacttaagctcaatac171b AflII R (SEQ ID NO: 478) gtattgagcttaagtcgggactg171b genomiq BsaI F  aaaGGTCTCCTAGC (SEQ ID NO: 479)TTGGTCAAACATACATACAGTAGCACTAG 171b genomiq BsaI R aaaGGTCTCgCGTAAAATTGTACTGATTG (SEQ ID NO: 480) AATGTAAATTGGTACACAthmiORF160b F (SEQ ID NO: 481) TAGCATGTTTTCCCCTCAATGAAthmiORF160b R (SEQ ID NO: 482) cgtaTCATTGAGGGGAAAACATAthMIR160b BsaI F (SEQ ID NO: 483) aaaGGTCTCcTAGCACTCATAACTCTCCCC AAATTCAthMIR160b BsaI R (SEQ ID NO: 484) aaaGGTCTCgCGTAGAAAGAATGTTCGAAAAACAATG pro 165a BsaI F (SEQ ID NO: 485)aGGTCTCCAAATcatgaagcaggcagtaataacct 165a muORF tagHA BsaI R aGGTCTCgCGTActaAGCGTAATCTGGAAC (SEQ ID NO: 486)ATCGTATGGGTAtaatatcctcgatccagacaac 165a fusion transcript BsaI R aGGTCTCgccatagtggcggagacgaagatg (SEQ ID NO: 487)165a fusion traduc BsaI R  aGGTCTCgccattaatatcctcgatccagacaac(SEQ ID NO: 488) pro 165a BsaI F (SEQ ID NO: 489)aGGTCTCCAAATtaaactgtcagtgcatggatgt 165a fusion GTG BsaI R aGGTCTCgACACattcacaaagttgtagagag (SEQ ID NO: 490)165a fusion GTGmutGTT BsaI aGGTCTCgACACattaacaaagttgtagagagR (SEQ ID NO: 491) 165a fusion CTG BsaI R aGGTCTCgACACtagcagattcacaaatttttgttg (SEQ ID NO: 492)165a fusion ATG BsaI R  aGGTCTCgACACcctcatgataatcgatcttagca(SEQ ID NO: 493)

B: Analysis of the miPEPs in Animals

Example 4 Identification of Candidate miPEPs in Drosophila

a) Identification of miPEP Candidates

A first study carried out by RACE-PCR in the model animal Drosophilamelanogaster shows the existence of miRNAs that are expressed duringembryogenesis, miR1 and miR8.

As in the plants, miORFs were identified in each of the two miRNAsstudied. For example, miR8, known for its role in the regulation ofgrowth in insects, has a miORF potentially encoding miPEP8.

Regarding DmmiR1 (identified in Drosophila melanogaster), it has twoDmmiORFs potentially encoding DmmiPEPla and DmmiPEP1b.

A phylogenetic analysis shows evolutionary conservation of the presenceof the miORFs among the dozen Drosophila species analysed, i.e. sincetheir divergence more than 60 million years ago (FIG. 13).

Moreover, the miPEPs identified in Drosophila have several similaritieswith the plant miPEPs. If their primary sequence and therefore theirsize evolve rapidly between species, a reduced size (from 32 to 104 AA)is found, as well as strong conservation for a basic overall charge (pHifrom 9.5 to 12) (FIG. 14).

Taken together, these results therefore indicate the existence ofregulatory miPEPs, encoded by the primary transcript of the microRNAs,over a broad spectrum of eukaryotic species. These discovered peptidesrepresent an as yet unexplored reservoir of natural molecules that mayregulate a variety of fundamental biological functions, both in plantsand in animals.

b) Expression of the miPEPs

Drosophila melanogaster cells have been transfected to overexpress theDmmiPEP8 (from the sequence SEQ ID NO: 494) or the DmmiPEP mt of whichstart codons have been mutated (from the sequence SEQ ID NO: 495). As atransfection control, one moesin-RFP-producing plasmid has beenco-transfected with the plasmids encoding DmmiPEP8 and DmmiPEP8 mt. Itis observed that start codons in the DmmiPEP8 sequence are functionalbecause they allow the synthesis of a miPEP::GFP fusion protein shown bythe GFP whereas the constructions with mutated ATGs do not producehardly any GFP.

These results suggest that the ORF of the miPEP is functional (FIG. 38).

Material and Methods

Cells of Drosophila melanogaster

S2 cells are cultured in a T75 flask in 12 mL of Schneider's medium(GIBCO), containing 1% of penicillin 100 U/mL and streptavidin 100 mg/mL(Sigma) and 10% of decomplemented foetal calf serum (30 min at 56° C.).

The transient transfections are carried out using the FuGENE® HDtransfection kit (Roche), according to the recommendations.Conventionally, 1.5 million S2 cells, previously seeded in 6-well plates(3 ml of medium per well), are transfected with 250 ng of total plasmidDNA. The DNA is brought into contact with the Fugene (3 μl) in 100 μl ofOPTIMEM (GIBCO). After 20 minutes, the transfection reagent formed isbrought into contact with the cells in the culture medium. The RNA ofthe cells is extracted 66 h after transfection.

Cloning of DmmiPEP8- and Mutated DmmiPEP8-Fragments

The DNA fragments, corresponding to the sequences of DmmiPEP8 (SEQ IDNO: 494) and of DmmiPEP8 mutated in the start codons (SEQ ID NO: 495),have been amplified by PCRin order to be cloned in accordance with theGFP in a pUAS plasmid. The SE insect cells have been co-transfected witha mix of plasmids (1/1/1 ratio, a total of 300 ng) encodingDmmiPEP8::GFP or its mutated form DmmiPEP8 mt::GFP, a pACT-GAL4 allowingtheir overexpression by means of a GAL4 transcription factor producedunder the control of the actin promotor and a expression vector encodingthe moesin-RFP protein used as a transfection control, also under thecontrol of the actin promotor. 48 h after transfection, the cells arefixed by the addition of formaldehyde in the final medium culture 4%(WN) for 10 min. Then, the cells are washed 2× with PBS 1X and coated onslides in order to be observed under a microscope (Leica SPE).

TABLE 9 Cloned sequences. CGCACACACACACACACACAATCACTCACACGCGGTCACACSEQ ID  GCACATTTCAATAAACTAATGGAGCCTGGCTTTGTTTTTGTT NO: 494TTATTTCCAACCCACTTGAGCACACAGCACACACAGAGAGAAAAATCAATACTCGTTATGGGATTAAATTTACAAAGCGCAAAGCAAAGCGACAAACAAAATTCAAAAGAAAGAAAAAAAAACACTCAAATAAACTCACAAAGAATTCCTTATCGCCAAGGGG GCCAATGTTCTAAGGTTCTTTCGCCTCGCACACACACACACACACAATCACTCACACGCGGTCACAC SEQ ID  GCACATTTCAATAAACTA AGTGAGCCTGGCTTTGTTTTTGTT NO: 495 TTATTTCCAACCCAC TTAAGCACACAGCACACACAGAGAGA AAAATCAATACTCGTT AGT GGATTAAATTTACAAAGCGCAAAGCAAAGCGACAAACAAAATTCAAAAGAAAGAAAAAAAAACACTCAAATAAACTCACAAAGAATTCCTTATCGCCAAGGGG GCCAATGTTCTAAGGTTCTTTCGCCT

C: Characterization of miPEPs in Humans

Example 5 Characterization of HsmiPEP155

The DNA fragments of interest (HsmiPEP155 and the mutated miPEP) weresynthesized or amplified by PCR using specific primers, and then clonedusing the enzymes XhoI and NotI into a pUAS plasmid permitting theiroverexpression by means of the GAL4 transcription factor, the expressionof which is controlled by a constitutive strong promoter.

The different constructs were produced either by PCR amplification ongenomic DNA of HeLa cells, or by RT-PCR on total RNAs of L428 humancells. The amplified PCR fragments are digested with the HindIII/EcoRIrestriction enzymes and then cloned into the vector pcDNA3.1. The DH5αstrain of Escherichia coli is electroporated and then cultured on asolid medium (2YT+agar+ampicillin). The plasmid DNA from differentclones is then prepared and sequenced for verification. The constructsare then prepared using the QIAfilter Plasmid Midi kit (QIAGEN) andstored at −20° C.

The HeLa cells (established tumour line, ATCC CCL-2.2) are cultured in a6-well plate in complete medium [(DMEM (1x)+Glutamax+4.5 g/L glucosewithout pyruvate+1x penicillin/streptomycin+1 mM Na-pyruvate+10% calfserum] and placed in an incubator at 37° C. and 5% CO₂.

The cells are transfected when they are at 50% confluence. At the startof the experiment, the complete medium containing the antibiotics isreplaced with complete medium without antibiotics.

For each well, a mix A [250 μl of Optimem (+Glutamax) (Gibco)+2 μg ofDNA] and a mix B [250 μl of Optimem+4 μl of Lipofectamine 2000(Invitrogen)] is prepared, and left for 5 min at ambient temperature.Then mix B is mixed dropwise into mix A, and left to incubate for 25 minat ambient temperature. The mixture is then deposited dropwise into thewell. 4-5 hours later, the medium is changed and replaced with completemedium with antibiotics. 48 hours after transfection, the cells arestopped. The medium is aspirated and discarded; the cells are rinsedwith PBS 1×. It is then possible to store the cells at −20° C. orextract the total RNAs directly.

For each well, the RNAs are extracted by depositing 1 ml of Tri-Reagent(Euromedex) on the cells. The Tri-reagent is aspirated and returnedseveral times so that the cells are lysed correctly, and then it istransferred into a 1.5-ml tube. 0.2 ml of water-saturated chloroform isadded. It is mixed by vortexing, then left for 2 to 3 minutes at ambienttemperature. It is centrifuged for 5 minutes at 15300 rpm and at 4° C.The aqueous phase is precipitated from 0.5 ml of isopropanol afterincubation for 10 minutes at ambient temperature and centrifugation for15 minutes at 15300 rpm and at 4° C. The supernatant is discarded andthe pellet is rinsed with 1 ml of 70% ethanol, with centrifugation for 5minutes at 15300 rpm at 4° C. The supernatant is again discarded and thepellet is dried for a few minutes in the air. For best-possible removalof the genomic DNA potentially remaining, the RNAs are treated withDNase. For this, the pellet is resuspended in 170 μl of ultra-purewater, 20 μl of DNase buffer 10× and 10 μl of RQ1 RNase-free DNase andheld at 37° C. for 30 minutes. Then 20 μl of SDS10% and 5 μl ofproteinase K (20 mg/ml) are added over 20 minutes at 37° C.

A last phenol extraction is carried out with 225 μl of aphenol/H₂O/chloroform mixture, and centrifuged for 5 minutes at 15300rpm at 4° C.

The aqueous phase is then precipitated from 20 μl of 3M sodium acetateand 600 μl of 100% ethanol for 20 minutes at −80° C. Then it iscentrifuged for 15 min at 4° C. at 15300 rpm. The supernatant isdiscarded. The pellet is rinsed in 1 ml of 70% ethanol, centrifuged for5 min at 15300 rpm at 4° C., the supernatant is discarded again and thepellet is left to dry for some minutes in the air.

The pellet is then taken up in 15-20 μl of ultra-pure water and the RNAsare assayed.

10-15 μg of total RNAs is then analysed by Northern blot on 15%acrylamide gel [solution of acrylamide/40% bis-acrylamide, ratio 19:1],7M urea in TBE 1x. Migration is carried out at 400V, in TBE1x asmigration buffer, after preheating the gel. The RNAs are thenelectro-transferred onto a Biodyne Plus 0.45 μm nylon membrane, for 2hours, at 1V and 4° C. in a transfer tank. At the end of transfer, themembrane is irradiated with UV at 0.124 J/cm². The membrane is thenpre-hybridized in a buffer 5× SSPE, 1× Denhardt's, 1% SDS and 150 μg/mlof yeast tRNA, for 1 hour at 50° C. in a hybridization oven. Then thenucleotide probe is added, labelled at 5′ with γ-³²P-ATP (0.5 to 1·10⁶cpm/ml of hybridization buffer) and is hybridized overnight at 50° C.The membrane is then washed twice in 0.1× SSPE/0.1% SDS at ambienttemperature and exposed in an autoradiography cassette containing aBioMax HE screen (Kodak) and a BioMax MS film (Kodak), in order todetect a microRNA, for 24-48 hours, at −80° C.

1. Process for detecting and identifying a micropeptide (miPEP) encodedby a nucleotide sequence contained in the sequence of the primarytranscript of a microRNA, comprising: a) a step of detecting an openreading frame from 12 to 303 nucleotides in length contained in thesequence of the primary transcript of said microRNA, then b) a step ofcomparison between: the accumulation of said microRNA in a specifiedeukaryotic cell expressing said microRNA, in the presence of a peptideencoded by a nucleotide sequence that is identical or degeneraterelative to that of said open reading frame, said peptide being presentin the cell independently of transcription of the primary transcript ofsaid microRNA, and the accumulation of said microRNA in a eukaryoticcell of the same type as the aforesaid specified eukaryotic cellexpressing said microRNA, in the absence of said peptide, in which amodulation of the accumulation of said microRNA in the presence of saidpeptide relative to the accumulation of said microRNA in the absence ofsaid peptide indicates the existence of a micropeptide encoded by saidopen reading frame.
 2. Process for detecting and identifying a miPEPaccording to claim 1, in which the modulation of the accumulation ofsaid microRNA is a decrease or an increase in the accumulation of saidmicroRNA, in particular an increase.
 3. Process for detecting andidentifying a miPEP according to claim 1, in which the presence of thepeptide in the cell results from: the introduction of a nucleic acidencoding said peptide into the cell, or the introduction of said peptideinto the cell.
 4. Process for detecting and identifying a miPEPaccording to claim 1, in which said open reading frame in step a) iscontained in the 5′ or 3′ portion of said primary transcript of themicroRNA, preferably in the 5′ portion.
 5. Process for detecting andidentifying a miPEP according to claim 1, in which said microRNA ispresent in a wild-type plant cell or in a wild-type animal cell, and inwhich said eukaryotic cell used in step b) is a plant cell, preferablyof a crucifer, of a leguminous plant or of a plant of the Solanaceaefamily or is an animal cell, preferably a human cell or Drosophila cell.6. Process for detecting and identifying a miPEP according to claim 1,in which said microRNA is of endogenous origin in said eukaryotic cellused in step b).
 7. Process for detecting and identifying a miPEPaccording to claim 1, in which said microRNA is of exogenous origin insaid eukaryotic cell used in step b), said eukaryotic cell containing avector allowing the expression of said microRNA.
 8. Process fordetecting and identifying a miPEP according to claim 1, in which theaccumulation of said microRNA is determined using quantitative RT-PCR,Northern blot, a DNA or RNA chip.
 9. MiPEP encoded by the nucleotidesequence as obtained by the process according to claim
 1. 10. MiPEP from3 to 100 amino acids, preferably from 4 to 40 amino acids, encoded by nopen reading frame from 12 to 303 nucleotids contained in a nucleotidesequence contained in the primary transcript of a microRNA, said miPEPbeing capable of modulating the accumulation of said microRNA in aeukaryotic cell, said nucleotide sequence being contained in the 5′ or3′ portion of said primary transcript of a microRNA, preferably in the5′ portion, the modulation of the microRNA by said miPEP being evidencedby observing a variation in the amounts of microRNA quantities betweencells in the presence or in the absence of miPEP.
 11. MiPEP according toclaim 9, said miPEP being selected from the group of peptides consistingof: SEQ ID NO: 1 to SEQ ID NO: 104 and SEQ ID NO: 355, SEQ ID NO: 375 toSEQ ID NO: 386, or SEQ ID NO:
 424. 12. Nucleic acid molecule encoding amiPEP as defined according to claim 9, said nucleic acid molecule beingselected in particular from the group of nucleic acids consisting of:SEQ ID NO: 105 to SEQ ID NO: 208 and SEQ ID NO: 356, SEQ ID NO: 387 toSEQ ID NO: 399, or SEQ ID NO:
 425. 13. A method for modulating theexpression of a gene in a specified eukaryotic cell, comprising exposingsaid specified eukaryotic cell at least: a miPEP as defined according toclaim 9, a nucleic acid encoding said miPEP, or a vector containing saidnucleic acid, said specified eukaryotic cell being capable of expressinga microRNA, the primary transcript of which contains at least onenucleotide sequence encoding said at least one miPEP and theaccumulation of which is modulated by said at least one miPEP, theexpression of said gene being regulated by said microRNA, said miPEP inparticular being selected from the group of peptides consisting of: SEQID NO: 1 to SEQ ID NO: 104 and SEQ ID NO: 355, SEQ ID NO: 375 to SEQ IDNO: 386, or SEQ ID NO: 424, in particular: SEQ ID NO: 59, SEQ ID NO: 24,SEQ ID NO: 43, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 14, SEQ ID NO:63 and SEQ ID NO: 424, said nucleic acid in particular being selectedfrom the group of nucleic acids consisting of: SEQ ID NO: 105 to SEQ IDNO: 208 and SEQ ID NO: 356, SEQ ID NO: 387 to SEQ ID NO: 399, or SEQ IDNO: 425, in particular: SEQ ID NO: 163, SEQ ID NO: 128, SEQ ID NO: 147,SEQ ID NO: 180, SEQ ID NO: 181, SEQ ID NO: 118, SEQ ID NO: 425 and SEQID NO: 167, said microRNA being selected in particular from the group ofnucleic acids consisting of: SEQ ID NO: 282 to SEQ ID NO: 354, and SEQID NO: 358, SEQ ID NO: 412 to SEQ ID NO: 423, or SEQ ID NO: 427, inparticular SEQ ID NO: 319, SEQ ID NO: 297, SEQ ID NO: 305, SEQ ID NO:331, SEQ ID NO: 291, SEQ ID NO: 322 and SEQ ID NO:
 427. 14. Compositioncomprising at least: a miPEP as defined according to claim 9, a nucleicacid encoding said miPEP, or a vector containing said nucleic acid. foruse as a human or veterinary medicament.
 15. Process for modulating theexpression of a gene regulated by a microRNA in a eukaryotic plant oranimal cell, in particular a human cell, comprising carrying out a stepof accumulation of a miPEP in said eukaryotic cell, said miPEP having: asize from 3 to 100 amino acids, preferably 4 to 20 amino acids, and apeptide sequence identical to that encoded by a nucleotide sequencecontained in the primary transcript of a microRNA regulating theexpression of said gene, and being capable of modulating theaccumulation of said microRNA, in which the accumulation of said miPEPin said eukaryotic cell induces a modulation of the expression of saidgene relative to the expression of said gene without accumulation ofsaid miPEP, said process not being used for surgical or therapeutictreatment of the human body or animal body, nor for modifying thegenetic identity of a human being.
 16. A method for promoting the growthand/or development of plants, in particular for modulating thephysiological parameters of a plant, in particular the biomass, foliarsurface area, flowering, fruit size, production and/or selection ofplant seeds, in particular for controlling the parthenocarpy ormonoecism of a plant, or for modifying the physiological parameters ofplant seeds, in particular germination, establishment of the root systemand resistance to water stress, or for preventing or treating plantdiseases,in particular for promoting resistance to infectious diseases,comprising administering to a plant in need there of a compositioncomprising at least: a miPEP as defined according to claim 9, a nucleicacid encoding said miPEP, or a vector containing said nucleic acid, as aphytopharmaceutical agent.
 17. A method for production and/or selectionof plant seeds, comprising applying a coating or film-coating of saidplant seeds a composition comprising at least: a miPEP as definedaccording claim 9, a nucleic acid encoding said miPEP, or a vectorcontaining said nucleic acid, as a phytopharmaceutical agent.
 18. Amethod for eradicating plants or slowing their growth comprisingadministering to the plants a composition at least: a miPEP as definedaccording to claim 9, a nucleic acid encoding said miPEP, or a vectorcontaining said nucleic acid, as a herbicide.
 19. A method foreradicating organisms harmful to plants or liable to be classified assuch, in particular as insecticide, arachnicide, molluscicide, orrodenticide, comprising applying to a plant or to a support in contactwith the plantcomposition comprising at least: a miPEP as definedaccording to claim 9, a nucleic acid encoding said miPEP, or a vectorcontaining said nucleic acid, as a pesticide.
 20. A compositioncomprising, in combination, a quantity of seeds of a plant and aquantity of a peptide the sequence of which comprises or consists of asequence identical to that of a miPEP naturally present in said plant,in particular said composition being formulated so as to form a coatedseed.